CN106962692A - A kind of premix for repairing the green liver pathological changes of grass carp and its preparation method and application - Google Patents
A kind of premix for repairing the green liver pathological changes of grass carp and its preparation method and application Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/60—Moraceae (Mulberry family), e.g. breadfruit or fig
- A61K36/605—Morus (mulberry)
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- A61K36/18—Magnoliophyta (angiosperms)
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- A61K36/46—Eucommiaceae (Eucommia family), e.g. hardy rubber tree
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
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- A61K36/65—Paeoniaceae (Peony family), e.g. Chinese peony
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/906—Zingiberaceae (Ginger family)
- A61K36/9066—Curcuma, e.g. common turmeric, East Indian arrowroot or mango ginger
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Abstract
The present invention relates to feed pre-mixing material, more particularly to a kind of premix for repairing the green liver pathological changes of grass carp, and the preparation method of the premix and its application, the premix is mainly made up of the component of following parts by weight：10 15 parts of mulberry leaf zymotic fluid, 10 25 parts of Fermented Liquor of Eucommia ulmoides Oliv Leaf, 15 parts of turmeric zymotic fluid, 40 60 parts of dregs of beans, 17 parts of root of herbaceous peony zymotic fluid.Compared with the conventional premix added in existing feed of grass carp, the present invention utilizes unique fermentation process, effective active composition in natural plants is set fully to discharge, there is significant adjustment effect to the dredging of the green liver pathological changes position bile duct of grass carp, the symptom of green liver pathological changes can be effectively repaired, the repair rate to the green liver pathological changes of grass carp can reach more than 90%.
Description
Technical field
The present invention relates to feed pre-mixing material, more particularly to a kind of premix for repairing the green liver pathological changes of grass carp, and the premix
Preparation method and its apply.
Background technology
Grass carp is the important economic freshwater breed variety of China, year cultured output account for the large freshwater aquiculture kind of China it
It is first.Liver of Ctenopharyngodon Idellus belongs to digestive system important component.Whether the function of Liver of Ctenopharyngodon Idellus health degree and digestive system reaches
Most preferably there is direct inner link, if liver health is impaired, each stage material ratio or feed conversion rate of grass carp all will be by
To bad influence.
Practice finds that annual lunar calendar solar term are clear and bright with before and after White Dew, often taking expelling parasite medication, sterilization medication, chemical drug
It it is also Liver of Ctenopharyngodon Idellus health problem the exposure obviously period, Liver of Ctenopharyngodon Idellus health problem Deng operation the most frequently period
One of them is exactly to contaminate the lesion being characterized so that liver part or major part appearance are green, is commonly called as green liver, shows as hepatic disease portion
The ruddy color and luster fading of hepatic tissue of position, and gradually by the gradual substitution of green.
The genesis mechanism of the green liver pathological changes of grass carp, be the factors such as medicine, constitution it is multiple under the influence of, be distributed in hepatic tissue it
Between bile duct occur different degrees of blocking, cause liver bile duct the cholestatic expansion of biliverdin occur, when liver bile duct parietal cell
When gap is loose, biliverdin permeates to hepatic tissue or spread into hepatic sinusoid to capillary network therewith, and liver is finally presented
The phenomenon of the green dye in local or big portion.
Generally, once occurring the green liver pathological changes of grass carp and continuing, if lacking effective recovery scenario, green liver is not only
Grass carp digestive function can be caused to decline, the decline of grass carp constitution premunition is also resulted in, grass carp old three disease is will also become when serious
Important risk factor.
The content of the invention
In a first aspect, the present invention provides a kind of premix for repairing the green liver pathological changes of grass carp, mainly by following parts by weight
Component is made：10-15 parts of mulberry leaf zymotic fluid, 10-25 parts of Fermented Liquor of Eucommia ulmoides Oliv Leaf, 1-5 parts of turmeric zymotic fluid is 40-60 parts of dregs of beans, white
1-7 parts of Chinese herbaceous peony zymotic fluid.
A kind of premix for repairing the green liver pathological changes of grass carp of the present invention is directed to above-mentioned pathogenesis or similar morbidity machine
The green liver pathological changes of the cultured fishes of reason have good repairing effect, especially for the green liver pathological changes of grass carp.
The present invention provides a kind of premix of the above-mentioned green liver pathological changes of reparation grass carp, wherein, mulberry leaf zymotic fluid, folium cortex eucommiae fermentation
Liquid, turmeric zymotic fluid and root of herbaceous peony zymotic fluid are the key function composition that green liver is repaired；Dregs of beans is the absorption carrier of liquid composition, hair
Wave filler function.
In the premix of the present invention, turmeric zymotic fluid and root of herbaceous peony zymotic fluid have good to phagocyte in grass carp hepatic tissue
Good activation, on the one hand can strengthen removing of the phagocyte to the downright bad liver cell in green liver pathological changes position；On the other hand can
Nutrient source is provided for grass carp hepatocyte regeneration, accelerates the self-regeneration and the elimination of green liver symptom of downright bad hepatic tissue；Fresh mulberry leaf is sent out
Closely connection has good repair to green liver pathological changes position bile duct cell wall for zymotic fluid and Fermented Liquor of Eucommia ulmoides Oliv Leaf, effectively alleviates
The problem of biliverdin exosmoses, while strengthening patency of the bile duct in bile transportation.
It is preferred that, a kind of premix for repairing the green liver pathological changes of grass carp is mainly prepared from by following parts by weight of component：Mulberry leaf
10-15 parts of zymotic fluid, 20-25 parts of Fermented Liquor of Eucommia ulmoides Oliv Leaf, 3-5 parts of turmeric zymotic fluid, 50-60 parts of dregs of beans, root of herbaceous peony zymotic fluid 5-7
Part.
In a preferred embodiment, a kind of premix for repairing the green liver pathological changes of grass carp, mainly by following parts by weight
Component is made：15 parts of mulberry leaf zymotic fluid, 25 parts of Fermented Liquor of Eucommia ulmoides Oliv Leaf, 5 parts of turmeric zymotic fluid, 50 parts of dregs of beans, root of herbaceous peony zymotic fluid 5
Part.
In another preferred embodiment, a kind of premix for repairing the green liver pathological changes of grass carp, mainly by following parts by weight
Component be made：10 parts of mulberry leaf zymotic fluid, 20 parts of Fermented Liquor of Eucommia ulmoides Oliv Leaf, 3 parts of turmeric zymotic fluid, 60 parts of dregs of beans, root of herbaceous peony zymotic fluid 7
Part.
It is preferred that, the mulberry leaf zymotic fluid is prepared by the method comprised the steps of：By mulberry leaf：Water：Gemma liquid
Culture medium and bacillus subtilis mixing, 37 DEG C of closed culture to OD values are that 3-4.5 obtains culture, and culture is centrifuged, received
The supernatant integrated is mulberry leaf zymotic fluid；Wherein, mulberry leaf (weight)：Water (volume)：The bulking value of gemma liquid culture medium (volume)
Than 1 (weight)：4-8 (volume)：1-3 (volume), the addition of bacillus subtilis is to add 1 ╳ per parts by weight mulberry leaf
105cfu-3╳105cfu。
Wherein, limit of the mixing without addition sequence of above-mentioned mulberry leaf, water, gemma liquid culture medium and bacillus subtilis
System, for example with first mulberry leaf, water and gemma liquid culture medium is mixed, adds bacillus subtilis.
Further, above-mentioned culture centrifugation is preferably centrifuged in 3000rpm, and such as 3000rpm is centrifuged 10 minutes.
In a preferred embodiment, a kind of mulberry leaf zymotic fluid, is prepared by the method comprised the steps of：By mulberry
Leaf：Water：Gemma liquid culture medium volume ratio 1 (weight) by weight：4-6 (volume)：1-1.5 (volume) is mixed to form mixed liquor,
Press again and add 1.5 ╳ 10 per parts by weight mulberry leaf5cfu-2.5╳105Cfu bacillus subtilises, 37 DEG C of closed culture to OD values are 3-
4.5, culture centrifugation is removed slag, the supernatant of collection is mulberry leaf zymotic fluid.
In a preferred embodiment, a kind of mulberry leaf zymotic fluid, is prepared by the method comprised the steps of：Will
The gemma liquid culture medium mixing of the mulberry leaf of 100 parts by weight, the water of 400 parts by volume and 100 parts by volume, then by every parts by weight mulberry leaf
Add 2 ╳ 105Cfu bacillus subtilises, 37 DEG C of constant-temperature enclosed culture to OD values are 3-4.5, and culture 3000rpm is centrifuged into 10
Minute removes slag, and it is mulberry leaf zymotic fluid to be collected into supernatant.
Wherein, the gemma liquid culture medium can be commercially available gemma liquid culture medium, the Bacillus of such as import
mediumbase。
Wherein, the bacillus subtilis is amphimicrobian type bacillus subtilis.
Wherein, the mulberry leaf are fresh mulberry leaf, the fresh mulberry leaf of such as water content >=80%, it is ensured that the freshness of mulberry leaf.
Wherein, the preferred sterile distilled water of water during prepared by above-mentioned mulberry leaf zymotic fluid.
It is preferred that, the Fermented Liquor of Eucommia ulmoides Oliv Leaf is prepared by the method comprised the steps of：By folium cortex eucommiae, water, gemma
Liquid culture medium and bacillus subtilis mixing, 37 DEG C of closed cultures obtain culture for 3.3-4.5 to OD values, by culture from
The heart removes slag, and the supernatant of collection is Fermented Liquor of Eucommia ulmoides Oliv Leaf；Wherein, the w/v of folium cortex eucommiae and water is 1：4-12, water and gemma
The volume ratio of liquid culture medium is 1-3：1, the addition of bacillus subtilis is to add 3 ╳ 10 per parts by weight folium cortex eucommiae3cfu-
4.5╳103cfu。
Wherein, limit of the mixing without addition sequence of above-mentioned folium cortex eucommiae, water, gemma liquid culture medium and bacillus subtilis
System, for example with first folium cortex eucommiae, water and gemma liquid culture medium is mixed, adds bacillus subtilis.
Further, above-mentioned culture centrifugation is preferably centrifuged in 3000rpm, and such as 3000rpm is centrifuged 10 minutes.
In a preferred embodiment, a kind of Fermented Liquor of Eucommia ulmoides Oliv Leaf, is prepared by the method comprised the steps of：Will
The volume ratio by weight 1 of folium cortex eucommiae and water：9 are mixed to form mixed liquor, by water and gemma liquid culture medium volume ratio 3：1 to mixing
Liquid adds gemma liquid culture medium, then adds 3.3 ╳ 10 by every parts by weight folium cortex eucommiae3Cfu bacillus subtilises, 37 DEG C of closed trainings
It is 3.8 to support to OD values, and culture 3000rpm is centrifuged 10 minutes, and the supernatant of collection is Fermented Liquor of Eucommia ulmoides Oliv Leaf.
In a preferred embodiment, a kind of Fermented Liquor of Eucommia ulmoides Oliv Leaf, is prepared by the method comprised the steps of：Will
The volume ratio by weight 1 of folium cortex eucommiae and water：6 are mixed to form mixed liquor, by anhydrous and gemma liquid culture medium volume ratio 2.6：1 to
Mixed liquor adds gemma liquid culture medium, then adds 3 ╳ 10 by every parts by weight folium cortex eucommiae3Cfu bacillus subtilises, 37 DEG C closed
Culture to OD values are 3.5, and culture 3000rpm is centrifuged 10 minutes, and the supernatant of collection is Fermented Liquor of Eucommia ulmoides Oliv Leaf.
Wherein, the folium cortex eucommiae is the folium cortex eucommiae of water content≤10%, and the control of water content is conducive to improving microbe conversion
Rate.
Wherein, the gemma liquid culture medium can be commercially available gemma liquid culture medium, the Bacillus of such as import
mediumbase。
Wherein, the bacillus subtilis is amphimicrobian type bacillus subtilis.
Wherein, the preferred sterile distilled water of water during prepared by above-mentioned Fermented Liquor of Eucommia ulmoides Oliv Leaf.
It is preferred that, the turmeric zymotic fluid is prepared by the method comprised the steps of：By turmeric, water, gemma liquid
Culture medium and bacillus subtilis mixing, 37 DEG C of closed culture to OD values are that 3-4.2 obtains culture, and culture is centrifuged, received
The supernatant integrated is turmeric zymotic fluid；Wherein, turmeric：Water：The w/v of gemma liquid culture medium is 1 (weight)：9-15 (bodies
Product)：3-5 (volume), the addition of bacillus subtilis is to add 1 ╳ 10 per parts by weight turmeric5cfu-4╳105cfu。
Wherein, limit of the mixing without addition sequence of above-mentioned turmeric, water, gemma liquid culture medium and bacillus subtilis
System, for example with first turmeric, water and gemma liquid culture medium is mixed, adds bacillus subtilis.
Further, above-mentioned culture centrifugation is preferably centrifuged in 3000rpm, and such as 3000rpm is centrifuged 10 minutes.
In a preferred embodiment, a kind of turmeric zymotic fluid, is prepared by the method comprised the steps of：Water：
Gemma liquid culture medium volume ratio 1 (weight) by weight：10-12 (volume)：3-4 (volume) is mixed, then by every parts by weight turmeric
Add 1 ╳ 105-3╳105Cfu bacillus subtilises, 37 DEG C of closed culture to OD values are that 3-4.2 obtains culture, by culture
3000rpm is centrifuged 10 minutes, and the supernatant of collection is turmeric zymotic fluid.
In a preferred embodiment, a kind of turmeric zymotic fluid, is prepared by the method comprised the steps of：Will
The gemma liquid culture medium mixing of the turmeric of 100 parts by weight, the water of 1000 parts by volume and 300 parts by volume, adds 105cfu/ml
The parts by volume of bacillus subtilis 300 above, 37 DEG C of closed culture to OD values are 3-4.2, and culture 3000rpm is centrifuged into 10 points
Clock removes slag, and it is turmeric zymotic fluid to be collected into supernatant.
Wherein, the gemma liquid culture medium can be commercially available gemma liquid culture medium, the Bacillus of such as import
mediumbase。
Wherein, the bacillus subtilis is amphimicrobian type bacillus subtilis.
Wherein, the turmeric is turmeric of the water content below 8%, to ensure the curcumin obtained compared with Gao get Liang.
Wherein, the preferred sterile distilled water of water during prepared by above-mentioned turmeric zymotic fluid.
It is preferred that, the root of herbaceous peony zymotic fluid is prepared by the method comprised the steps of：By the root of herbaceous peony, water, gemma liquid
Culture medium and bacillus subtilis mixing, 37 DEG C of closed culture to OD values are that 3.5-4.8 obtains culture, and culture is centrifuged,
The supernatant of collection is root of herbaceous peony zymotic fluid；Wherein, the root of herbaceous peony：Water：The w/v of gemma liquid culture is 1 (weight)：3-8 (bodies
Product)：0.5-5 (volume), the addition of bacillus subtilis is to add 1 ╳ 10 per the parts by weight root of herbaceous peony6cfu-4╳106cfu。
Wherein, limit of the mixing without addition sequence of the above-mentioned root of herbaceous peony, water, gemma liquid culture medium and bacillus subtilis
System, for example with first by the mixing of the root of herbaceous peony, water and gemma liquid culture medium, then bacillus subtilis.
Further, above-mentioned culture centrifugation is preferably centrifuged in 3000rpm, and such as 3000rpm is centrifuged 10 minutes.
In a preferred embodiment, a kind of root of herbaceous peony zymotic fluid, is prepared by the method comprised the steps of：Will be white
Chinese herbaceous peony：Water：Gemma liquid culture medium volume ratio 1 (weight) by weight：3-5 (volume)：0.5-2 (volume) is mixed, then by every weight
Part root of herbaceous peony adds 1 ╳ 106cfu-3╳106Cfu bacillus subtilises, 37 DEG C of closed culture to OD values are that 3.5-4.2 is cultivated
Thing, culture 3000rpm is centrifuged 10 minutes, and the supernatant of collection is root of herbaceous peony zymotic fluid.
In a preferred embodiment, a kind of root of herbaceous peony zymotic fluid, is prepared by the method comprised the steps of：Will
The gemma liquid culture medium mixing of the root of herbaceous peony of 100 parts by weight, the water of 300 parts by volume and 50 parts by volume, then add per the parts by weight root of herbaceous peony
Enter 106More than the cfu/ml parts by volume of bacillus subtilis 100,37 DEG C it is constant-temperature enclosed culture 96 hours, by culture 3000rpm
Centrifugation is removed slag for 10 minutes, and it is root of herbaceous peony zymotic fluid to be collected into supernatant.
Wherein, the gemma liquid culture medium can be commercially available gemma liquid culture medium, the Bacillus of such as import
mediumbase。
Wherein, the bacillus subtilis is amphimicrobian type bacillus subtilis.
Wherein, the root of herbaceous peony is the root of herbaceous peony of the water content below 8%.
Wherein, the preferred sterile distilled water of water during prepared by above-mentioned root of herbaceous peony zymotic fluid.
The premix of the present invention is using unique viable bacteria fermentation technology, to these four plants of mulberry leaf, folium cortex eucommiae, turmeric and the root of herbaceous peony
Thing carries out biofermentation, and the tunning of acquisition is rich in various active peptide, amino acid, vitamin, trace element, to green liver pathological changes
The dredging of position bile duct has significant adjustment effect, can effectively repair the symptom of green liver pathological changes；To fish body and breeding environment
Residual is not produced, and there is green safety environmental protection.
In second aspect, the present invention provides a kind of method for the premix for preparing the above-mentioned green liver pathological changes of reparation grass carp, described
Method comprises the following steps：The component is mixed to get mixture, is dried at 30-40 DEG C to the moisture of mixture and is
8-10%, and be alternatively crushed in 80-120 mesh sieves, obtain premix.
In the embodiment of the preferred premix for preparing the above-mentioned green liver pathological changes of reparation grass carp, this method includes following
Step：The component is mixed to get mixture, it is 8-10%, and powder to be dried at 30-35 DEG C to the moisture of mixture
It is broken in 100-120 mesh sieves, obtains premix.
In the embodiment of the preferred premix for preparing the above-mentioned green liver pathological changes of reparation grass carp, this method includes following
Step：The component is mixed to get mixture, it is 10% to be dried at 30 DEG C to the moisture of mixture, and is crushed to
In 120 mesh sieves, premix is obtained.
In the third aspect, the present invention also provides a kind of application of the premix of the above-mentioned green liver pathological changes of reparation grass carp, for pre-
Anti- property repairs the green liver pathological changes of grass carp and/or for the therapeutic reparation green liver pathological changes of grass carp.
Wherein, when the premix is used for preventative reparation grass carp green liver pathological changes, 1-1.5% makes an addition to by weight percentage
In feed, preferably continuous was a cycle using 7-15 days.
Wherein, when the premix is used for therapeutic reparation grass carp green liver pathological changes, the additions of 1.5-2.5% by weight percentage
In feed, preferably continuous was a cycle using 7-15 days.
Compared with the conventional premix added in existing feed of grass carp, the present invention makes day using unique fermentation process
Effective active composition fully discharges in right plant, has significant adjustment effect to the dredging of the green liver pathological changes position bile duct of grass carp,
The symptom of green liver pathological changes can be effectively repaired, the repair rate to the green liver pathological changes of grass carp can reach more than 90%.
Embodiment
The present invention is further described below in conjunction with specific embodiment.
Embodiment 1
A kind of premix for repairing the green liver pathological changes of grass carp, is prepared by the component of following parts by weight：Mulberry leaf zymotic fluid 15
Part, 25 parts of Fermented Liquor of Eucommia ulmoides Oliv Leaf, 5 parts of turmeric zymotic fluid, 50 parts of dregs of beans, 5 parts of root of herbaceous peony zymotic fluid.
Wherein, mulberry leaf zymotic fluid is prepared by the method for following steps：By mulberry leaf：Sterile distilled water：Gemma liquid is trained
Foster base volume ratio 1 (weight) by weight：4-6 (volume)：1-1.5 (volume) is mixed to form mixed liquor, then by every parts by weight mulberry leaf
Add 1.5 ╳ 105cfu-2.5╳105Cfu bacillus subtilises, 37 DEG C of closed culture to OD values are 3-4.5, and culture is centrifuged
Remove slag, the supernatant of collection is mulberry leaf zymotic fluid.
Wherein, Fermented Liquor of Eucommia ulmoides Oliv Leaf is prepared by the method for following steps：Folium cortex eucommiae and pressing for sterile distilled water are weighed
Measure volume ratio 1：6 are mixed to form mixed liquor, by sterile distilled water and gemma liquid culture medium volume ratio 2.6：1 adds to mixed liquor
Gemma liquid culture medium, then add 3 ╳ 10 by every parts by weight folium cortex eucommiae3Cfu bacillus subtilises, 37 DEG C of closed cultures to OD values
For 3.5, culture 3000rpm is centrifuged 10 minutes, the supernatant of collection is Fermented Liquor of Eucommia ulmoides Oliv Leaf.
Wherein, turmeric zymotic fluid is prepared by the method for following steps：By the turmeric of 100 parts by weight, 1000 parts by volume
Water and 300 parts by weight gemma liquid culture medium mixing, add 105More than the cfu/ml volume of bacillus subtilis 300
Part, 37 DEG C of closed culture to OD values are that 3-4.2 obtains culture, and culture 3000rpm centrifugations are removed slag for 10 minutes, are collected into
It is turmeric zymotic fluid clearly.
Wherein, root of herbaceous peony zymotic fluid is prepared by the method for following steps：By the root of herbaceous peony：Sterile distilled water：Gemma liquid is trained
Foster base volume ratio 1 (weight) by weight：3-5 (volume)：0.5-2 (volume) is mixed, then adds 1 ╳ by every parts by weight root of herbaceous peony
106cfu-3╳106Cfu bacillus subtilises, 37 DEG C of closed culture to OD values are that 3.5-4.2 obtains culture, by culture
3000rpm is centrifuged 10 minutes, and the supernatant of collection is root of herbaceous peony zymotic fluid.
The method for preparing the premix is：Each component is sufficiently mixed, 30 DEG C of lasting drying are then placed in mixing
During thing water content≤10%, mixture is crossed into 120 mesh pulverizers and crushed, the crushed material of acquisition is premix.
Embodiment 2
A kind of premix for repairing the green liver pathological changes of grass carp, is prepared by the component of following parts by weight：Mulberry leaf zymotic fluid 10
Part, 20 parts of Fermented Liquor of Eucommia ulmoides Oliv Leaf, 3 parts of turmeric zymotic fluid, 60 parts of dregs of beans, 7 parts of root of herbaceous peony zymotic fluid.
Wherein, mulberry leaf zymotic fluid is prepared by the method for following steps：By the mulberry leaf of 100 parts by weight, 400 parts by volume
Water and the mixing of the gemma liquid culture medium of 100 parts by volume, then add 2 ╳ 10 by every parts by weight mulberry leaf5Cfu bacillus subtilises,
37 DEG C of constant-temperature enclosed culture to OD values are 3-4.5, and culture 3000rpm centrifugations are removed slag for 10 minutes, supernatant as mulberry is collected into
Leaf zymotic fluid.
Wherein, Fermented Liquor of Eucommia ulmoides Oliv Leaf is prepared by the method for following steps：Folium cortex eucommiae and pressing for sterile distilled water are weighed
Measure volume ratio 1：9 are mixed to form mixed liquor, by sterile distilled water and gemma liquid culture medium volume ratio 3：1 adds bud to mixed liquor
Spore liquid culture medium, then add 3.3 ╳ 10 by every parts by weight folium cortex eucommiae3Cfu bacillus subtilises, 37 DEG C of closed cultures to OD values
For 3.8, culture 3000rpm is centrifuged 10 minutes, the supernatant of collection is Fermented Liquor of Eucommia ulmoides Oliv Leaf.
Wherein, turmeric zymotic fluid is prepared by the method for following steps：Sterile distilled water：Gemma liquid culture medium is by weight
Measure volume ratio 1 (weight)：10-12 (volume)：3-4 (volume) is mixed, then adds 1 ╳ 10 by every parts by weight turmeric5-3╳105cfu
Bacillus subtilis, 37 DEG C of closed culture to OD values are that 3-4.2 obtains culture, and culture 3000rpm is centrifuged 10 minutes, received
The supernatant integrated is turmeric zymotic fluid.
Wherein, root of herbaceous peony zymotic fluid, is prepared by the method for following steps：By the root of herbaceous peony of 100 parts by weight, 300 parts by volume
Water and 50 parts by volume the mixing of gemma liquid culture medium, then add per the parts by weight root of herbaceous peonys 106More than cfu/ml withered grass gemma
The parts by volume of bacillus 100,37 DEG C it is constant-temperature enclosed culture 96 hours, by culture 3000rpm centrifugation remove slag within 10 minutes, be collected into supernatant
As root of herbaceous peony zymotic fluid.
The method for preparing the premix is：Each component is sufficiently mixed, 30 DEG C of lasting drying are then placed in mixing
During thing water content≤10%, mixture is crossed into 120 mesh pulverizers and crushed, the crushed material of acquisition is premix.
The premix of the present invention of test example 1 is assessed the preventative repairing effect of the green liver pathological changes of grass carp
Commercially available grass carp expanded pellet feed product, as the basal feed of this experimental example, the nutrition of basal feed product into
Divide main：Albumen 30%, fat 8%, crude fibre 12%, coarse ash 16%, lysine 2.8%.
(1) premix material additive amount group is set
The carrier based on commercial diet product, adds premix prepared by different amounts of embodiment 1 to carrier is carrier respectively
The group of material is as test group, while mulberry leaf zymotic fluid, Fermented Liquor of Eucommia ulmoides Oliv Leaf, turmeric zymotic fluid and the root of herbaceous peony in setting premix respectively
Premix when zymotic fluid is individually lacked is added as parallel control group by not same amount, wherein using without premix as blank pair
According to group.Test group sets details to be shown in Table 1：
Table 1：Different tests group configuration information table
Note：It is formulated based on the premix formulation that corresponding scarce composition premix formulation is prepared by embodiment 1 in group,
Scarce composition proportional quantity with etc. weight flour be replaced.
(2) experiment packet and management
63g ± 5g healthy grass carp is selected, 4800 tails is selected at random from 5500 tail grass carps, is randomly divided into 16 groups, wherein
Every group divide into 3 it is parallel, each group feeds above-mentioned 16 kinds of feeds respectively.24 days test periods.During experiment, each group grass carp presses 2-
2.5% average daily feeding rate is fed, and three meals in a day right place is fed.
(3) the preventative repairing effect of green liver pathological changes is assessed
When experiment the 1-7 days, each group continuously fed corresponding testing material.Test the 8-12 days and raise side for oral administration by mixing
Method, to each test group feed in feed by 35mg/kg fish bodies weight it is daily, by the praziquantel and commercial diet of effective content 2%
It is sufficiently mixed, preservation of drying in the shade feeds successive administration by being satiated with food 5 days, stops administration.Test continuous three days the 13-15 days and do not feed intake,
Hungry stress is formed to cultivation grass carp.Above-mentioned test operation is to be based on production experience, simulates multifactor superposition and induces grass carp
Green liver pathological changes.Test the 16-23 days, feed the corresponding testing material of each test group within continuous 8 days.When testing the 24th day, to each
Test group, which is synchronized, to be analysed, and exposure liver is simultaneously taken pictures, if the form of the slight green dye of locality occurs in liver, is regarded as slight
Green liver pathological changes；If the form of the more dark green dye of locality occurs in liver, the green liver pathological changes of moderate are regarded as；If liver occur big portion or
All the form of green dye, then regard as the green liver pathological changes of severe.According to how much statistics of the green liver quantity of grass carp during each group off-test
Green liver pathological changes incidence and repair rate, its calculation formula are as follows.
Green liver pathological changes incidence (%)=identification meets grass carp quantity/same group experiment grass carp Zong Shuo Liang ╳ of green liver pathological changes
100
When testing the 24th day, each group grass carp is synchronized to analyse live confirmation, with control group grass carp slight green liver pathological changes, moderate
On the basis of green liver pathological changes and the green liver pathological changes incidence of severe, the different degrees of green liver pathological changes incidence of each group grass carp of statistics and repair
Multiple rate such as table 2.
Table 2：
From table 2 it can be seen that when there is no the premix of the addition present invention in the commercially available basal feed as blank control,
Endo-medicine and the green liver pathological changes incidence of grass carp that hungry stress induces are different, slight, moderate and the green hepatopathy variation of severe
54%, 4%, 3% is not reached.When in basal feed add embodiment 1 prepare premix when, first, 0.5%, 1.0%,
1.5% 3 additive capacity group grass carp is slight, moderate and the preventative repair rate of the green liver pathological changes of severe and premix material additive amount height
It is low to be proportionate；Secondly, compared with blank control group, during premix addition 0.5% of the present invention, to the green liver pathological changes incidence of grass carp
With good preventative repairing effect, and when premix material additive amount increases to 1.5%, its prevention to the green liver pathological changes of grass carp
Property repairing effect is more notable；Thus illustrate, with embodiment 1 prepare premix in the range of recommended dose dosage it is upper
Rise, the preventative repair rate of the different degrees of green liver pathological changes of high additive capacity group grass carp is respectively provided with good using effect；With blank
Green liver pathological changes incidence is reference to control group grass carp in various degree, and the use of this premix is preventative to slight green liver pathological changes to repair
Rate is 72-96%, and the preventative repair rate highest of the green liver pathological changes of alignment degree and severe is respectively 100% and 100%.
When in basal feed respectively addition missing mulberry leaf zymotic fluid, missing Fermented Liquor of Eucommia ulmoides Oliv Leaf, missing turmeric zymotic fluid and
When lacking the premix of the present invention of root of herbaceous peony zymotic fluid, correspondence component missing 0.5%, 1.0% and 1.5% 3 dosage group grass carp
Green liver repair rate is proportionate with additive capacity, i.e., with the rising of additive capacity, and green liver incidence declines change in correlation,
And the green liver incidence of three dosage group grass carps is substantially compared with the reduction of blank control group, but its incidence is still significantly higher than correspondence component
The premix of the present invention of zero missing.Thus illustrate, when correspondence component missing in premix of the present invention, although added by adjusting
Dosage can improve the preventative repairing effect of green liver of the premix of correspondence component missing, but its preventative repairing effect is still reached to
Less than green liver repairing effect of the premix of the present invention to application dosage.It can be seen that, mulberry leaf zymotic fluid, Fermented Liquor of Eucommia ulmoides Oliv Leaf, ginger
Yellow zymotic fluid and root of herbaceous peony zymotic fluid any component have to other compositions in the present invention in the green liver pathological changes of preventative reparation grass carp
Synergistic function, also, mulberry leaf zymotic fluid, Fermented Liquor of Eucommia ulmoides Oliv Leaf, turmeric zymotic fluid and root of herbaceous peony zymotic fluid are preventative reparations
The key function composition of the green liver pathological changes of grass carp.
The premix of the present invention of test example 2 is assessed the therapeutic repairing effect of the green liver pathological changes of grass carp
Commercially available grass carp expanded pellet feed product, as the basal feed of this experimental example, the nutrition of basal feed product into
Divide predominantly：Albumen 30%, fat 8%, crude fibre 12%, coarse ash 16%, lysine 2.8%.
(1) premix material additive amount group is set
The carrier based on commercial diet product, adds premix prepared by different amounts of embodiment 2 to carrier respectively
Group is as test group, while mulberry leaf zymotic fluid, Fermented Liquor of Eucommia ulmoides Oliv Leaf, turmeric zymotic fluid and root of herbaceous peony fermentation in setting premix respectively
Premix when liquid is individually lacked is added as parallel control group by not same amount, wherein being used as blank using the group without premix
Control group.Test group sets details to be shown in Table 3：
Table 3：Different tests group configuration information table
Note：It is formulated based on the premix formulation that corresponding scarce composition premix formulation is prepared by embodiment 2 in group,
Scarce composition proportional quantity with etc. weight flour be replaced.
(2) experiment packet and management
63g ± 5g healthy grass carp is selected, 4800 tails is selected at random from 5500 tail grass carps, is randomly divided into 16 groups, wherein
Every group divide into 3 it is parallel, each group feeds above-mentioned 16 kinds of feeds respectively.22 days test periods.During experiment, each group grass carp is pressed
2.0-2.5% average daily feeding rate is fed, and three meals in a day right place is fed.
(3) structure of green liver pathological changes pathological model
Based on production experience, simulate multifactor superposition and induce the green liver pathological changes of grass carp.When experiment the 1-8 days, continuous 8
It does not feed feed, and hungry stress is formed to cultivation grass carp.Test the 9-13 days, in the way of mixing feeding and take orally, by effective content
2% praziquantel is sufficiently mixed with commercial diet and pharmaceutical chemistry is made, it is ensured that medicine intake dosage reaches 35mg/kg fishes in bait of offeing medicine
Body weight is daily.Test continuous 8 days the 14-21 days and feed the corresponding testing material of each test group.
(4) the therapeutic repairing effect of green liver pathological changes is assessed
When testing the 22nd day, each test group is synchronized and analysed, exposure liver is simultaneously taken pictures, if locality occurs in liver
The form of slight green dye, then regard as slight green liver pathological changes；If there is the form of the more dark green dye of locality in liver, in regarding as
Spend green liver pathological changes；If the form of the green dye of most or all occurs in liver, the green liver pathological changes of severe are regarded as.Tested and tied according to each group
The green liver pathological changes incidence of how much statistics and repair rate of the green liver quantity of grass carp during beam, its calculation formula are as follows.
Green liver pathological changes incidence (%)=identification meets grass carp quantity/same group experiment grass carp Zong Shuo Liang ╳ of green liver pathological changes
100
When testing the 22nd day, each group grass carp is synchronized to analyse live confirmation, the different degrees of green liver pathological changes hair of each group grass carp
Raw rate and repair rate such as table 4.
Table 4：
From table 4, it can be seen that when there is no the premix of the addition present invention in the commercially available basal feed as blank control,
Endo-medicine and the green liver pathological changes incidence of grass carp that hungry stress induces are different, slight, moderate and the green hepatopathy variation of severe
64%, 5%, 7% is not reached.When in basal feed add embodiment 2 prepare premix when, first, 1.5%, 2.0%,
2.5% 3 additive capacity group grass carp is slight, moderate and the therapeutic repair rate of the green liver pathological changes of severe and premix material additive amount height
It is low to be proportionate；Secondly, compared with blank control group, during premix addition 1.5% of the present invention, to the green liver pathological changes incidence of grass carp
With good therapeutic repairing effect, and when premix material additive amount increases to 2.5%, its treatment to the green liver pathological changes of grass carp
Property repairing effect is more notable；Thus illustrate, with embodiment 2 prepare premix in the range of recommended dose dosage it is upper
Rise, the therapeutic repair rate of the different degrees of green liver pathological changes of high additive capacity group grass carp is respectively provided with good using effect；With blank
Green liver pathological changes incidence is reference to control group grass carp in various degree, and the use of this premix is to slight green liver pathological changes repair rate
The therapeutic repair rate highest of green liver pathological changes of 61-94%, alignment degree and severe is respectively 100% and 100%.
When in basal feed respectively addition missing mulberry leaf zymotic fluid, missing Fermented Liquor of Eucommia ulmoides Oliv Leaf, missing turmeric zymotic fluid and
When lacking the premix of the present invention of root of herbaceous peony zymotic fluid, correspondence component missing 1.5%, 2.0% and 2.5% 3 dosage group grass carp
Green liver repair rate is proportionate with additive capacity, i.e., with the rising of additive capacity, and green liver incidence declines change in correlation,
And the green liver incidence of three dosage group grass carps is substantially compared with the reduction of blank control group, but its incidence is still significantly higher than correspondence component
The premix of the present invention of zero missing.Thus illustrate, when correspondence component missing in premix of the present invention, although added by adjusting
Dosage can improve the therapeutic repairing effect of green liver of the premix of correspondence component missing, but its therapeutic repairing effect is still reached to
Less than green liver therapeutic repairing effect of the premix of the present invention to application dosage.It can be seen that, mulberry leaf zymotic fluid, folium cortex eucommiae fermentation
Liquid, turmeric zymotic fluid and root of herbaceous peony zymotic fluid any component are to other compositions in the present invention in the green liver pathological changes of therapeutic reparation grass carp
With synergistic function, also, mulberry leaf zymotic fluid, Fermented Liquor of Eucommia ulmoides Oliv Leaf, turmeric zymotic fluid and root of herbaceous peony zymotic fluid are therapeutic
Repair the key function composition of the green liver pathological changes of grass carp.
It is described above, it is only the preferably specific embodiment of the present invention, but protection scope of the present invention is not limited thereto,
Any one skilled in the art the invention discloses technical scope in, technique according to the invention scheme and its
Design is subject to equivalent substitution or change, should all cover within the scope of the present invention.
Claims (10)
1. a kind of premix for repairing the green liver pathological changes of grass carp, it is characterised in that be mainly made up of the component of following parts by weight：Mulberry leaf
10-15 parts of zymotic fluid, 10-25 parts of Fermented Liquor of Eucommia ulmoides Oliv Leaf, 1-5 parts of turmeric zymotic fluid, 40-60 parts of dregs of beans, root of herbaceous peony zymotic fluid 1-7
Part.
2. a kind of premix for repairing the green liver pathological changes of grass carp according to claim 1, it is characterised in that mainly by following heavy
The component of amount part is made：10-15 parts of mulberry leaf zymotic fluid, 20-25 parts of Fermented Liquor of Eucommia ulmoides Oliv Leaf, 3-5 parts of turmeric zymotic fluid, dregs of beans 50-
60 parts, 5-7 parts of root of herbaceous peony zymotic fluid.
3. a kind of premix for repairing the green liver pathological changes of grass carp according to claim 2, it is characterised in that mainly by following heavy
The component of amount part is made：15 parts of mulberry leaf zymotic fluid, 25 parts of Fermented Liquor of Eucommia ulmoides Oliv Leaf, 5 parts of turmeric zymotic fluid, 50 parts of dregs of beans, root of herbaceous peony hair
5 parts of zymotic fluid.
4. a kind of premix for repairing the green liver pathological changes of grass carp according to claim 2, it is characterised in that mainly by following heavy
The component of amount part is made：10 parts of mulberry leaf zymotic fluid, 20 parts of Fermented Liquor of Eucommia ulmoides Oliv Leaf, 3 parts of turmeric zymotic fluid, 60 parts of dregs of beans, root of herbaceous peony hair
7 parts of zymotic fluid.
5. according to a kind of premix of any described green liver pathological changes of reparation grass carp in Claims 1-4, it is characterised in that institute
Mulberry leaf zymotic fluid is stated to be prepared by the method comprised the steps of：By mulberry leaf：Water：Gemma liquid culture medium and bacillus subtilis
Bacterium mixes, and 37 DEG C of closed culture to OD values are that 3-4.5 obtains culture, and culture is centrifuged, and the supernatant of collection ferments for mulberry leaf
Liquid；Wherein, mulberry leaf：Water：The w/v 1 of gemma liquid culture medium：4-8：1-3, the addition of bacillus subtilis is every
Parts by weight mulberry leaf add 1 ╳ 105cfu-3╳105cfu。
6. according to a kind of premix of any described green liver pathological changes of reparation grass carp in Claims 1-4, it is characterised in that institute
Fermented Liquor of Eucommia ulmoides Oliv Leaf is stated to be prepared by the method comprised the steps of：By folium cortex eucommiae, water, gemma liquid culture medium and withered grass bud
Spore bacillus mixes, and 37 DEG C of closed culture to OD values are that 3.3-4.5 obtains culture, and culture centrifugation is removed slag, the supernatant of collection
For Fermented Liquor of Eucommia ulmoides Oliv Leaf；Wherein, the w/v of folium cortex eucommiae and water is 1：The volume ratio of 4-12, water and gemma liquid culture medium
For 1-3：1, the addition of bacillus subtilis is to add 3 ╳ 10 per parts by weight folium cortex eucommiae3cfu-4.5╳103cfu。
7. according to a kind of premix of any described green liver pathological changes of reparation grass carp in Claims 1-4, it is characterised in that institute
Turmeric zymotic fluid is stated to be prepared by the method comprised the steps of：By turmeric, water, gemma liquid culture medium and bacillus subtilis
Bacterium mixes, and 37 DEG C of closed culture to OD values are that 3-4.2 obtains culture, and culture is centrifuged, and the supernatant of collection ferments for turmeric
Liquid；Wherein, turmeric：Water：The w/v of gemma liquid culture medium is 1：9-15：3-5, the addition of bacillus subtilis is
1 ╳ 10 is added per parts by weight turmeric5cfu-4╳105cfu。
8. according to a kind of premix of any described green liver pathological changes of reparation grass carp in Claims 1-4, it is characterised in that institute
Root of herbaceous peony zymotic fluid is stated to be prepared by the method comprised the steps of：By the root of herbaceous peony, water, gemma liquid culture medium and bacillus subtilis
Bacterium mixes, and 37 DEG C of closed culture to OD values are that 3.5-4.8 obtains culture, and culture is centrifuged, and the supernatant of collection is sent out for the root of herbaceous peony
Zymotic fluid；Wherein, the root of herbaceous peony：Water：The w/v of gemma liquid culture is 1：3-8：0.5-5, the addition of bacillus subtilis
1 ╳ 10 is added for every parts by weight root of herbaceous peony6cfu-4╳106cfu。
9. a kind of method prepared according to the premix of any described reparation green liver pathological changes of grass carp in claim 1 to 8, it is special
Levy and be, the described method comprises the following steps：The component is mixed to get mixture, dried at 30-40 DEG C to mixture
Moisture be 8-10%, and be alternatively crushed in 80-120 mesh sieves, obtain premix.
10. a kind of method applied according to the premix of any described reparation green liver pathological changes of grass carp in claim 1 to 8, its
It is characterised by, the premix is used for the preventative reparation green liver pathological changes of grass carp and/or for the therapeutic reparation green liver pathological changes of grass carp.
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