CN108850610B - Composite plant extract feed additive and preparation method and application thereof - Google Patents

Composite plant extract feed additive and preparation method and application thereof Download PDF

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CN108850610B
CN108850610B CN201810864535.1A CN201810864535A CN108850610B CN 108850610 B CN108850610 B CN 108850610B CN 201810864535 A CN201810864535 A CN 201810864535A CN 108850610 B CN108850610 B CN 108850610B
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weight
parts
feed additive
plant extract
composite plant
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CN108850610A (en
Inventor
李同刚
周盛昌
肖俊峰
张军
刘小成
傅心锋
杨再龙
张敬学
吴有林
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Hunan Aonong Biotechnology Co ltd
Jinhua Aonong Biological Science & Technology Co ltd
Fujian Aonong Biological Technology Group Co Ltd
Zhangzhou Aonong Animal Husbandry Technology Co Ltd
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Hunan Aonong Biotechnology Co ltd
Jinhua Aonong Biological Science & Technology Co ltd
Fujian Aonong Biological Technology Group Co Ltd
Zhangzhou Aonong Animal Husbandry Technology Co Ltd
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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K50/00Feeding-stuffs specially adapted for particular animals
    • A23K50/30Feeding-stuffs specially adapted for particular animals for swines
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K10/00Animal feeding-stuffs
    • A23K10/10Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/14Pretreatment of feeding-stuffs with enzymes
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K10/00Animal feeding-stuffs
    • A23K10/30Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K50/00Feeding-stuffs specially adapted for particular animals
    • A23K50/60Feeding-stuffs specially adapted for particular animals for weanlings

Abstract

The invention relates to a composite plant extract feed additive and a preparation method and application thereof, belonging to the field of feeds. The raw materials comprise purslane, stevia rebaudiana leaves, acanthopanax, astragalus, Chinese violet and callicarpa. The feed additive has reasonable formula, natural components, no toxic or side effect and no drug resistance, is beneficial to regulating the organism immunity of live pigs, promotes the growth of the live pigs and reduces the morbidity. The preparation method comprises the following steps: mixing the above raw materials at a certain ratio to obtain a primary material, mixing the primary material with water and a complex enzyme, performing enzymolysis, concentrating, and drying. The preparation method is simple to operate, is rapid and efficient, and can enable the feed additive to have a good effect. The product can be used for preparing pig feed, and can significantly improve feed intake and immunity of pig, and reduce diarrhea.

Description

Composite plant extract feed additive and preparation method and application thereof
Technical Field
The invention relates to the field of feeds, and in particular relates to a composite plant extract feed additive and a preparation method and application thereof.
Background
For years, the feed antibiotics play an important role in the aspects of preventing and treating livestock and poultry diseases, promoting growth and the like. However, abuse of antibiotics causes serious bacterial drug resistance and drug residue, causes various problems of food safety, environmental pollution and the like, and seriously threatens the health and safety of human beings.
Therefore, the research of adding a safe, green, novel and efficient antibiotic substitute into the feed becomes a hot spot of the research.
Disclosure of Invention
The invention aims to provide a composite plant extract feed additive which has a reasonable formula, natural components, no toxic or side effect and no drug resistance, is beneficial to regulating the organism immunity of live pigs, promotes the growth of the live pigs and reduces the morbidity.
The second purpose of the invention is to provide a preparation method of the composite plant extract feed additive, which is simple to operate, fast and efficient, and can enable the feed additive to have better action effect.
The third purpose of the invention is to provide an application of the composite plant extract feed additive, which can obviously improve the feed intake and immunity of live pigs and reduce diarrhea when being used for preparing live pig feed.
The technical problem to be solved by the invention is realized by adopting the following technical scheme:
the invention provides a composite plant extract feed additive, which comprises, by weight, 11-12 parts of purslane, 6-8 parts of stevia rebaudiana leaves, 3-5 parts of acanthopanax, 1-3 parts of astragalus membranaceus, 1-2 parts of Chinese violet and 4-6 parts of callicarpa bodinieri.
The invention also provides a preparation method of the composite plant extract feed additive, which comprises the following steps: mixing herba Portulacae, folium Chrysanthemi, radix Acanthopanacis Senticosi, radix astragali, herba Violae and folium Callicarpae Formosanae at a certain ratio to obtain primary material, mixing the primary material with water and complex enzyme, performing enzymolysis, concentrating, and drying.
The complex enzyme comprises cellulase and glucose oxidase.
Preferably, the weight ratio of cellulase to glucose oxidase is 1: 0.5-0.8.
The invention also provides application of the composite plant extract feed additive, for example, the composite plant extract feed additive can be used for preparing live pig feed.
Preferably, the compound plant extract feed additive is added into the live pig feed according to the proportion of 0.01-0.1 wt%.
More preferably, the compound plant extract feed additive is added into the live pig feed according to the proportion of 0.05 wt%.
The composite plant extract feed additive provided by the preferred embodiment of the invention and the preparation method and application thereof have the beneficial effects that:
the compound plant extract feed additive provided by the application has the advantages of reasonable formula, natural components, no toxic or side effect and no drug resistance, and is beneficial to regulating the organism immunity of live pigs, promoting the growth of the live pigs and reducing the morbidity. The preparation method is simple to operate, is rapid and efficient, and can enable the feed additive to have a good effect. The product can be used for preparing pig feed, and can significantly improve feed intake and immunity of pig, and reduce diarrhea.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
The composite plant extract feed additive, the preparation method and the application thereof according to the embodiment of the present invention are specifically described below.
The raw materials of the composite plant extract feed additive provided by the invention comprise 11-12 parts by weight of purslane, 6-8 parts by weight of stevia rebaudiana leaves, 3-5 parts by weight of acanthopanax, 1-3 parts by weight of astragalus mongholicus, 1-2 parts by weight of philippine violet herb and 4-6 parts by weight of callicarpa bodinieri.
In some embodiments, the above raw materials may include 11.2-11.8 parts by weight of purslane, 6.5-7.5 parts by weight of stevia rebaudiana leaves, 3.5-4.5 parts by weight of acanthopanax, 1.5-2.5 parts by weight of astragalus membranaceus, 1.2-1.8 parts by weight of viola yedoensis and 4.5-5.5 parts by weight of callicarpa bodinieri.
In some particularly preferred embodiments, the above raw materials may include 11.5 parts by weight of purslane, 7 parts by weight of stevia leaves, 4 parts by weight of acanthopanax, 2 parts by weight of astragalus, 1.5 parts by weight of viola yedoensis and 5 parts by weight of callicarpa bodinieri.
Wherein the herba Portulacae has effects of removing toxic substance, relieving inflammation, promoting diuresis, and relieving swelling, and can effectively reduce levels of Bacillus dysenteriae, Bacillus typhi, Escherichia coli and Staphylococcus aureus.
Stevia rebaudiana leaves contain sweet components stevioside with low calorie and high sweetness, so that the stevia rebaudiana leaves can help digestion and promote functions of pancreas and spleen and stomach.
Acanthopanax senticosus contains a large amount of bioactive factors and phenolic substances, and can enhance the immunity of the organism, improve the internal circulation environment and balance the metabolism of glycolipid, and the superoxide dismutase SOD complex contained in the acanthopanax senticosus has good oxidation resistance. In addition, the acanthopanax contains rich vitamins, minerals and various amino acid components, can well supplement the nutrition of the feed, and improves the production performance of the live pigs while enhancing the immunity.
After the purslane, the stevia rebaudian leaves and the acanthopanax are mixed, the effects of strengthening the middle-jiao and replenishing qi, strengthening the lung and benefiting the spleen, diminishing inflammation and promoting urination and the like can be achieved, the level of harmful bacteria in live pigs, such as dysentery bacillus, typhoid bacillus, escherichia coli and staphylococcus aureus, can be reduced, and the intestinal flora regulating effect is good. In addition, the palatability of the product can be improved and the feed intake of the live pigs can be improved after the substances are matched.
The Chinese violet contains various flavonoid compounds, coumarin compounds, phenolic compounds and the like, has the effects of clearing heat and removing toxicity, cooling blood and relieving swelling, clearing heat and promoting diuresis, can effectively treat diseases such as jaundice, dysentery and diarrhea, enhances the immunocompetence of live pigs, and improves the digestion and absorption capacity of the live pigs.
The Callicarpa Nudiflora contains polysaccharide, flavonoid, condensed tannin, resveratrol, stevioside, vanillin and other nutritional components, and can effectively inhibit Escherichia coli, Shigella flexneri, Staphylococcus aureus, streptococcus and the like. After the callicarpa bodinieri and the purslane are matched, the effect of reducing harmful flora can be further strengthened, and the intestinal health of the live pigs is protected.
The astragalus root which has the functions of promoting the metabolism of the live pig organism, promoting the renewal of serum and liver proteins and enhancing and regulating the immunity of the organism is added into the formula, so that the comprehensive immunity of the live pig can be further improved, various harmful pathogenic bacteria causing diarrhea can be inhibited and killed, the intestinal health of the live pig can be effectively improved, and the rapid growth can be promoted.
In conclusion, the composite plant extract feed additive has the advantages of green and natural components, no toxic or side effect, no drug resistance, contribution to regulating the organism immunity of live pigs, promotion of the growth of the live pigs, reduction of the morbidity and capability of replacing antibiotics in the prior art.
In addition, the application also provides a preparation method of the composite plant extract feed additive, which comprises the following steps: mixing herba Portulacae, folium Chrysanthemi, radix Acanthopanacis Senticosi, radix astragali, herba Violae and folium Callicarpae Formosanae at a certain ratio to obtain primary material, mixing the primary material with water and complex enzyme, performing enzymolysis, concentrating, and drying.
For reference, the feed-to-liquid ratio of the initial material to the water may be 1 g: 5-10mL, e.g., 1 g: 5mL, 1 g: 6mL, 1 g: 7mL, 1 g: 8mL, 1 g: 9mL or 1 g: 10mL, etc.
For reference, the weight ratio of the primary material to the complex enzyme may be 100: 0.1-0.5, e.g. 100: 0.1, 100: 0.2, 100: 0.3, 100: 0.4 or 100: 0.5, etc.
In the present application, the complex enzyme includes cellulase and glucose oxidase.
On one hand, the cellulase is used for decomposing cellulose of plant raw materials into protein of oligosaccharide or monosaccharide and destroying the integrity of plant cells, so that active substances contained in the plants are dissolved out of the cells; on the other hand, the digestibility of neutral detergent fiber and acidic detergent fiber in the live pig body can be improved.
Since cellulases from different sources have a wide difference in structure and function, cellulases produced by fungi such as Trichoderma, Aspergillus or Penicillium can be used as the cellulase used in the present application, and cellulases produced by bacteria can also be used.
In some specific embodiments, the Cellulase is Cellulase R-10(Cellulase R-10) which is purchased from Shanghai-sourced bioscience, Inc. and is numbered E0332, and by contrast, the effect of the feed additive prepared by the preparation method of the application on live pigs, particularly piglets, by using the Cellulase is obviously better than that of other cellulases.
The glucose oxidase can catalyze the enzymolysis of saccharides contained in plants, and the other can consume molecular oxygen or atomic oxygen to oxidize glucose, so that the easily-oxidized active ingredients (particularly phenolic compounds) in the plant active ingredients released by the enzymolysis of the cellulase are protected from being oxidized, and the phenomenon that the easily-oxidized ingredients generate quinone compounds after being oxidized and play a role in inhibiting the enzymolysis of the cellulase is avoided.
In some specific embodiments, the Glucose oxidase is Glucose oxidase (Glucose oxidase) which is purchased from Shanghai Biotechnology limited and is numbered E0750, and by contrast, the action effect of the feed additive prepared by the preparation method of the application by using the Glucose oxidase is obviously better than that of other Glucose oxidases on live pigs, especially piglets.
It should be noted that the mode of obtaining cellulase and glucose oxidase referred to in the present application is not limited to the above description, and can be obtained from other normal routes according to actual conditions.
In the present application, the weight ratio of the cellulase to the glucose oxidase may be, for example, 1: 0.5-0.8. The matching effect of the two enzymes is best under the mixture ratio.
In some embodiments, the enzymatic hydrolysis may be carried out at 45-65 ℃ for 8-12 hours to achieve a preferred level of enzymatic hydrolysis of both cellulase and glucose oxidase.
In some preferred embodiments, during the enzymatic hydrolysis, cysteine may also be added to the enzymatic hydrolysis system, which may be used in combination with glucose oxidase to further eliminate the formation of quinone compounds after oxidation of easily-oxidizable active ingredients (such as phenolic compounds) decomposed from plant materials under the action of cellulase, thereby inhibiting the enzymatic hydrolysis of cellulase. On the other hand, after entering the live pig body, the glutathione can be generated, not only can be combined with free radicals or heavy metals and the like in the live pig body to participate in the biotransformation function, but also can convert harmful toxicants in the live pig body into harmless substances to be discharged out of the body; but also can coordinate with vitamins in the live pigs to protect the health level of the pigs.
In some embodiments, the weight ratio of cysteine to cellulase may be 0.2 to 0.4: 1, e.g. 0.2: 1. 0.25: 1. 0.3: 1. 0.35: 1 or 0.4: 1.
in this application, the concentration and drying may be carried out by transferring the enzymatic hydrolysate (including the enzymatic hydrolysate and the precipitate) to a freeze-drying bottle, concentrating at 60-80 deg.C (for example, rotary evaporation to a thick state), and freeze-drying at-55 deg.C for 12-24h to obtain powdered composite plant extract feed additive.
Furthermore, before mixing with water and the complex enzyme, the method also comprises the steps of sequentially shearing, drying and crushing the primary material, and then sieving the primary material by a 30-50-mesh sieve, so that the enzymolysis effect and the product yield are improved.
According to the preparation method, the yield of the composite plant extract feed additive is 90-95 wt%. It is worth noting that the yield is in percent by weight of the final feed additive based on the weight of the raw materials.
In addition, the application also provides an application of the composite plant extract feed additive, for example, the composite plant extract feed additive can be used for preparing live pig feed.
In some embodiments, the composite plant extract feed additive is added to the live pig feed in a ratio of 0.01-0.1 wt%. In some preferred embodiments, the composite plant extract feed additive is added into the live pig feed in a proportion of 0.05 wt%, and has high cost performance.
The features and properties of the present invention are described in further detail below with reference to examples.
Example 1
The embodiment provides a preparation method of a composite plant extract feed additive, which comprises the following steps: mixing 11 parts by weight of purslane, 8 parts by weight of stevia rebaudiana leaves, 3 parts by weight of acanthopanax, 3 parts by weight of astragalus membranaceus, 1 part by weight of Chinese violet and 6 parts by weight of callicarpa bodinieri, shearing, drying, crushing, sieving by a 30-mesh sieve, and repeating the crushing and sieving steps until large particles all pass through 30 meshes to obtain a primary material.
And (3) mixing the primary material according to the material-liquid ratio of 1 g: mixing 10mL of the mixture with deionized water, stirring the mixture evenly, adding complex enzyme, and performing enzymolysis for 8 hours at 65 ℃ and under the condition of natural pH. The weight ratio of the primary material to the complex enzyme is 100: 0.5, the compound enzyme is prepared from the following components in percentage by weight of 1: 0.5 of cellulase and glucose oxidase. The cellulase is an Allantin brand cellulase with product number C128647 derived from Trichoderma reesei ATCC 26921. The glucose oxidase is prepared from Aspergillus niger with product number of G109029.
Transferring the enzymolysis product into a freeze drying bottle, performing rotary evaporation at 80 ℃ to obtain a thick product, and performing freeze drying at-55 ℃ for 24 hours to obtain the composite plant extract feed additive, wherein the yield is 95%.
Example 2
The embodiment provides a preparation method of a composite plant extract feed additive, which comprises the following steps: mixing 11 parts by weight of purslane, 8 parts by weight of stevia rebaudiana leaves, 3 parts by weight of acanthopanax, 3 parts by weight of astragalus membranaceus, 1 part by weight of Chinese violet and 6 parts by weight of callicarpa bodinieri, shearing, drying, crushing, sieving by a 40-mesh sieve, and repeating the crushing and sieving steps until large particles all pass through 40 meshes to obtain a primary material.
And (3) mixing the primary material according to the material-liquid ratio of 1 g: mixing 7mL of the mixture with deionized water, stirring uniformly, adding a complex enzyme, and performing enzymolysis for 10 hours at 55 ℃ and under the condition of natural pH. The weight ratio of the primary material to the complex enzyme is 100: 0.3, the compound enzyme is prepared from the following components in percentage by weight of 1: 0.8 of cellulase and glucose oxidase. The cellulase is an Allantin brand cellulase with product number C128647 derived from Trichoderma reesei ATCC 26921. The glucose oxidase is prepared from Aspergillus niger with product number of G109029.
Transferring the enzymolysis product into a freeze drying bottle, performing rotary evaporation at 70 ℃ to obtain a thick product, and performing freeze drying at-55 ℃ for 18h to obtain the composite plant extract feed additive, wherein the yield is 93%.
Example 3
The embodiment provides a preparation method of a composite plant extract feed additive, which comprises the following steps: mixing 11 parts by weight of purslane, 8 parts by weight of stevia rebaudiana leaves, 3 parts by weight of acanthopanax, 3 parts by weight of astragalus membranaceus, 1 part by weight of Chinese violet and 6 parts by weight of callicarpa bodinieri, shearing, drying, crushing, sieving by a 50-mesh sieve, and repeating the crushing and sieving steps until large particles all pass through 50 meshes to obtain a primary material.
And (3) mixing the primary material according to the material-liquid ratio of 1 g: 5mL of the mixture is mixed with deionized water, the mixture is stirred uniformly, and then complex enzyme is added for enzymolysis for 12 hours at the temperature of 45 ℃ and under the condition of natural pH. The weight ratio of the primary material to the complex enzyme is 100: 0.1, the compound enzyme is prepared from the following components in percentage by weight of 1: 0.65 cellulase and glucose oxidase. The cellulase is an Allantin brand cellulase with product number C128647 derived from Trichoderma reesei ATCC 26921. The glucose oxidase is prepared from Aspergillus niger with product number of G109029.
Transferring the enzymolysis product into a freeze drying bottle, performing rotary evaporation at 60 ℃ to obtain a thick product, and performing freeze drying at-55 ℃ for 12h to obtain the composite plant extract feed additive, wherein the yield is 92%.
Example 4
The embodiment provides a preparation method of a composite plant extract feed additive, which comprises the following steps: mixing 11.5 parts by weight of purslane, 7 parts by weight of stevia rebaudiana leaves, 4 parts by weight of acanthopanax, 2 parts by weight of astragalus membranaceus, 1.5 parts by weight of Chinese violet and 5 parts by weight of callicarpa bodinieri, shearing, drying, crushing, sieving with a 30-mesh sieve, and repeating the crushing and sieving steps until all large particles pass through 30 meshes to obtain a primary material.
And (3) mixing the primary material according to the material-liquid ratio of 1 g: mixing 10mL of the mixture with deionized water, stirring the mixture evenly, adding complex enzyme, and performing enzymolysis for 8 hours at 65 ℃ and under the condition of natural pH. The weight ratio of the primary material to the complex enzyme is 100: 0.5, the compound enzyme is prepared from the following components in percentage by weight of 1: 0.5 of cellulase and glucose oxidase. The cellulase is an Allantin brand cellulase with product number C128647 derived from Trichoderma reesei ATCC 26921. The glucose oxidase is prepared from Aspergillus niger with product number of G109029.
Transferring the enzymolysis product into a freeze drying bottle, performing rotary evaporation at 80 ℃ to obtain a thick product, and performing freeze drying at-55 ℃ for 24 hours to obtain the composite plant extract feed additive, wherein the yield is 94%.
Example 5
The embodiment provides a preparation method of a composite plant extract feed additive, which comprises the following steps: mixing 11.5 parts by weight of purslane, 7 parts by weight of stevia rebaudiana leaves, 4 parts by weight of acanthopanax, 2 parts by weight of astragalus membranaceus, 1.5 parts by weight of Chinese violet and 5 parts by weight of callicarpa bodinieri, shearing, drying, crushing, sieving with a 40-mesh sieve, and repeating the crushing and sieving steps until all large particles pass through 40 meshes to obtain a primary material.
And (3) mixing the primary material according to the material-liquid ratio of 1 g: mixing 7mL of the mixture with deionized water, stirring uniformly, adding a complex enzyme, and performing enzymolysis for 10 hours at 55 ℃ and under the condition of natural pH. The weight ratio of the primary material to the complex enzyme is 100: 0.3, the compound enzyme is prepared from the following components in percentage by weight of 1: 0.8 of cellulase and glucose oxidase. The cellulase is an Allantin brand cellulase with product number C128647 derived from Trichoderma reesei ATCC 26921. The glucose oxidase is prepared from Aspergillus niger with product number of G109029.
Transferring the enzymolysis product into a freeze drying bottle, performing rotary evaporation at 70 ℃ to obtain a thick product, and performing freeze drying at-55 ℃ for 18h to obtain the composite plant extract feed additive, wherein the yield is 93%.
Example 6
The embodiment provides a preparation method of a composite plant extract feed additive, which comprises the following steps: mixing 11.5 parts by weight of purslane, 7 parts by weight of stevia rebaudiana leaves, 4 parts by weight of acanthopanax, 2 parts by weight of astragalus membranaceus, 1.5 parts by weight of Chinese violet and 5 parts by weight of callicarpa bodinieri, shearing, drying, crushing, sieving with a 50-mesh sieve, and repeating the crushing and sieving steps until all large particles pass through 50 meshes to obtain a primary material.
And (3) mixing the primary material according to the material-liquid ratio of 1 g: 5mL of the mixture is mixed with deionized water, the mixture is stirred uniformly, and then complex enzyme is added for enzymolysis for 12 hours at the temperature of 45 ℃ and under the condition of natural pH. The weight ratio of the primary material to the complex enzyme is 100: 0.1, the compound enzyme is prepared from the following components in percentage by weight of 1: 0.65 cellulase and glucose oxidase. The cellulase is an Allantin brand cellulase with product number C128647 derived from Trichoderma reesei ATCC 26921. The glucose oxidase is prepared from Aspergillus niger with product number of G109029.
Transferring the enzymolysis product into a freeze drying bottle, performing rotary evaporation at 60 ℃ to obtain a thick product, and performing freeze drying at-55 ℃ for 12h to obtain the composite plant extract feed additive, wherein the yield is 93%.
Example 7
The embodiment provides a preparation method of a composite plant extract feed additive, which comprises the following steps: mixing 12 parts by weight of purslane, 6 parts by weight of stevia rebaudiana leaves, 5 parts by weight of acanthopanax, 1 part by weight of astragalus membranaceus, 2 parts by weight of Chinese violet and 4 parts by weight of callicarpa bodinieri, shearing, drying, crushing, sieving by a 30-mesh sieve, and repeating the crushing and sieving steps until large particles all pass through 30 meshes to obtain a primary material.
And (3) mixing the primary material according to the material-liquid ratio of 1 g: mixing 10mL of the mixture with deionized water, stirring the mixture evenly, adding complex enzyme, and performing enzymolysis for 8 hours at 65 ℃ and under the condition of natural pH. The weight ratio of the primary material to the complex enzyme is 100: 0.5, the compound enzyme is prepared from the following components in percentage by weight of 1: 0.5 of cellulase and glucose oxidase. The cellulase is an Allantin brand cellulase with product number C128647 derived from Trichoderma reesei ATCC 26921. The glucose oxidase is prepared from Aspergillus niger with product number of G109029.
Transferring the enzymolysis product into a freeze drying bottle, performing rotary evaporation at 80 ℃ to obtain a thick product, and performing freeze drying at-55 ℃ for 24 hours to obtain the composite plant extract feed additive, wherein the yield is 93%.
Example 8
The embodiment provides a preparation method of a composite plant extract feed additive, which comprises the following steps: mixing 12 parts by weight of purslane, 6 parts by weight of stevia rebaudiana leaves, 5 parts by weight of acanthopanax, 1 part by weight of astragalus membranaceus, 2 parts by weight of Chinese violet and 4 parts by weight of callicarpa bodinieri, shearing, drying, crushing, sieving by a 40-mesh sieve, and repeating the crushing and sieving steps until large particles all pass through 40 meshes to obtain a primary material.
And (3) mixing the primary material according to the material-liquid ratio of 1 g: mixing 7mL of the mixture with deionized water, stirring uniformly, adding a complex enzyme, and performing enzymolysis for 10 hours at 55 ℃ and under the condition of natural pH. The weight ratio of the primary material to the complex enzyme is 100: 0.3, the compound enzyme is prepared from the following components in percentage by weight of 1: 0.8 of cellulase and glucose oxidase. The cellulase is an Allantin brand cellulase with product number C128647 derived from Trichoderma reesei ATCC 26921. The glucose oxidase is prepared from Aspergillus niger with product number of G109029.
Transferring the enzymolysis product into a freeze drying bottle, performing rotary evaporation at 70 ℃ to obtain a thick product, and performing freeze drying at-55 ℃ for 18h to obtain the composite plant extract feed additive, wherein the yield is 91%.
Example 9
The embodiment provides a preparation method of a composite plant extract feed additive, which comprises the following steps: mixing 12 parts by weight of purslane, 6 parts by weight of stevia rebaudiana leaves, 5 parts by weight of acanthopanax, 1 part by weight of astragalus membranaceus, 2 parts by weight of Chinese violet and 4 parts by weight of callicarpa bodinieri, shearing, drying, crushing, sieving by a 50-mesh sieve, and repeating the crushing and sieving steps until large particles all pass through 50 meshes to obtain a primary material.
And (3) mixing the primary material according to the material-liquid ratio of 1 g: 5mL of the mixture is mixed with deionized water, the mixture is stirred uniformly, and then complex enzyme is added for enzymolysis for 12 hours at the temperature of 45 ℃ and under the condition of natural pH. The weight ratio of the primary material to the complex enzyme is 100: 0.1, the compound enzyme is prepared from the following components in percentage by weight of 1: 0.65 cellulase and glucose oxidase. The cellulase is an Allantin brand cellulase with product number C128647 derived from Trichoderma reesei ATCC 26921. The glucose oxidase is prepared from Aspergillus niger with product number of G109029.
Transferring the enzymolysis product into a freeze drying bottle, performing rotary evaporation at 60 ℃ to obtain a thick product, and performing freeze drying at-55 ℃ for 12h to obtain the composite plant extract feed additive, wherein the yield is 90%.
Example 10
This example differs from example 2 in that: the raw materials comprise 11.2 parts by weight of purslane, 6.5 parts by weight of stevia rebaudiana leaves, 3.5 parts by weight of acanthopanax, 1.5 parts by weight of astragalus membranaceus, 1.2 parts by weight of Chinese violet and 4.5 parts by weight of callicarpa bodinieri.
Example 11
This example differs from example 2 in that: the raw materials comprise 11.8 parts by weight of purslane, 7.5 parts by weight of stevia rebaudiana leaves, 4.5 parts by weight of acanthopanax, 2.5 parts by weight of astragalus membranaceus, 1.8 parts by weight of Chinese violet and 5.5 parts by weight of callicarpa bodinieri.
Example 12
This example differs from example 2 in that: the Cellulase is Cellulase R-10(Cellulase R-10) with the number of E0332 from Shanghai-derived bioscience GmbH.
Example 13
This example differs from example 2 in that: the Glucose oxidase is Glucose oxidase (Glucose oxidase) of Shanghai-derived Biotechnology Co., Ltd, No. E0750.
Example 14
This example differs from example 2 in that: the Cellulase is Cellulase R-10(Cellulase R-10) with the number of E0332 of Shanghai-derived Polybioscience, Inc., and the Glucose oxidase is Glucose oxidase (Glucose oxidase) with the number of E0750 of Shanghai-derived Polybioscience, Inc.
Example 15
This example differs from example 2 in that: during the enzymolysis process, cysteine is added into an enzymolysis system. The weight ratio of cysteine to cellulase is 0.2: 1.
example 16
This example differs from example 2 in that: during the enzymolysis process, cysteine is added into an enzymolysis system. The weight ratio of cysteine to cellulase is 0.3: 1.
example 17
This example differs from example 2 in that: during the enzymolysis process, cysteine is added into an enzymolysis system. The weight ratio of cysteine to cellulase is 0.4: 1.
example 18
The embodiment provides an application of a composite plant extract feed additive, namely adding the composite plant extract feed additive into live pig feed according to the proportion of 0.01 wt%. The compound plant extract feed additive can be prepared according to any one of the above examples 1-17.
Example 19
The embodiment provides an application of a composite plant extract feed additive, namely adding the composite plant extract feed additive into live pig feed according to the proportion of 0.1 wt%. The compound plant extract feed additive can be prepared according to any one of the above examples 1-17.
Example 20
The embodiment provides an application of a composite plant extract feed additive, namely adding the composite plant extract feed additive into live pig feed according to the proportion of 0.05 wt%. The compound plant extract feed additive can be prepared according to any one of the above examples 1-17.
Test example 1
The above examples 1-17 were repeated to obtain sufficient composite plant extract feed additives.
The following tests were carried out by taking examples 2, 6 and 7 as examples. The average weight of 150 healthy piglets in a breeding base of the Suzhou breeding base of Fujianhao agricultural biotechnology group Limited is about 18kg, and the weight difference is not obvious through statistical analysis. Randomly divided into 5 groups of 3 replicates each, with 10 replicates each. The treatment groups are respectively as follows: a blank control group, an antibiotic group and a test group, wherein the test group is 3 groups, the compound plant extract feed additives prepared in the examples 2, 6 and 7 are respectively added, and the adding proportion of the compound plant extract feed additives in the live pig feed is 0.05%. The specific test design is shown in table 1.
The piglets can freely take food and drink water, and are immunized and repelled according to the conventional method. Before the formal test, the piglet is firstly subjected to a one-week pre-feeding test in a period of 30 days, and then the piglet is subjected to material setting and weighing, and the daily gain, the feed intake, the material weight ratio and the diarrhea rate are calculated; at the end of the test, randomly selecting 3 test pigs in each repetition, fasting for 12h, taking 10mL of blood from the anterior vena cava, placing the blood obliquely for 30min at room temperature, centrifuging to obtain a serum sample, freezing for inspection, and measuring biochemical indexes.
The test results are shown in tables 2 to 4.
TABLE 1 design of the experiments
TABLE 2 Effect of different test groups on piglet growth Performance
TABLE 3 Effect of different test groups on piglet diarrhea Rate
TABLE 4 Effect of different test groups on the serum immune index of piglets
As can be seen from table 2, compared with the blank control group and the antibiotic group, the feed added with the composite plant extract feed additive prepared in the embodiments 2, 6 and 7 of the present invention has the advantages that the average daily gain and the average daily feed intake of piglets are improved after eating, and the feed-weight ratio is reduced, which indicates that the growth performance of piglets can be improved by feeding the live pig feed containing the composite plant extract feed additive provided in the embodiments of the present invention.
In the test group A, the test group B and the test group C, the test group A has the best effect, and the average daily gain is improved by 18.85 percent compared with a blank control group and is improved by 12.14 percent compared with an antibiotic group; the average daily feed intake is improved by 5.16 percent compared with a blank control group and is improved by 4.28 percent compared with an antibiotic group; the material weight is reduced by 12.05 percent compared with that of a blank control group and 7.01 percent compared with that of an antibiotic group.
As can be seen from table 3, compared with the blank control group, the feed added with the composite plant extract feed additive prepared in the embodiments 2, 6 and 7 of the present invention has a significantly reduced diarrhea rate after being eaten by piglets, which indicates that the disease resistance of live pigs can be improved and the diarrhea incidence can be reduced by feeding live pig feed containing the composite plant extract feed additive provided in the embodiments of the present invention. Compared with the antibiotic group, the diarrhea rate difference is not large, which indicates that the composite plant extract feed additive provided by the application can be used for replacing antibiotics.
In combination with table 4, it can be seen that, compared with the blank control group, the feed added with the composite plant extract feed additive prepared in the embodiments 2, 6 and 7 of the present invention has obviously improved immunoglobulin IgG, IgM and IgA in the serum of piglets after eating; compared to the antibiotic group, IgG, IgM were slightly higher in test group A, but IgA was slightly lower in test group A than in the antibiotic group. Results of interleukin-2 (IL-2) and gamma-interferon (IFN-gamma) in serum show that compared with a blank control group and an antibiotic group, the feed added with the feed additive prepared in the embodiments 2, 6 and 7 of the invention has the advantages that IL-2 and IFN-gamma are improved after piglets eat the feed, and the result shows that the immunity of piglet organisms can be improved by feeding the piglet feed containing the compound plant extract feed additive provided by the embodiment of the invention.
Comparing the test group A, the test group B and the test group C, wherein the indexes of the test group A, such as daily gain, material weight ratio, diarrhea rate, serum immunity index and the like, are better, and the optimal proportion and preparation process of the raw materials in the test group are shown.
In addition, the results of the tests on the feeds respectively containing the composite plant extract feed additives obtained in the other examples according to the methods of the test examples also show that the daily gain and daily feed intake of piglets can be generally increased, the feed-weight ratio can be reduced, the serum immunity index can be improved, and the diarrhea rate can be reduced compared with the blank control group and the antibiotic group by feeding the piglet feed added with the composite plant extract feed additive provided by the invention example.
Test example 2
Taking example 2 and examples 12-14 as examples, comparative groups 1-4 were set, wherein the Cellulase in comparative group 1 was Cellulase (Trichoderma viride) numbered E0330 from shanghai-sourced biosciences ltd (celluloase, Trichoderma viride G); the glucose oxidase of the comparative group 2 is the glucose oxidase with the product number of G130084, wherein the brands of the arabidopsis is derived from Aspergillus niger; comparative group 3 differs from example 2 in that: the cellulase part is replaced by equal amount of glucose oxidase, and the original glucose oxidase is not changed; comparative group 4 differs from example 2 in that: the glucose oxidase part is replaced by the same amount of cellulase, and the original cellulase is not changed.
The composite plant extract feed additive of the above 8 enzymes was added to the feed for piglets in the same amount according to the test method of test example 1 and fed to piglets. The results are shown in tables 5 to 7.
TABLE 5 Effect of different test groups on piglet growth Performance
TABLE 6 Effect of different test groups on piglet diarrhea Rate
TABLE 7 Effect of different test groups on the serum immune index of piglets
It can be seen from tables 5 to 7 that the effects on the growth performance, the immune function and the diarrhea rate of piglets of the comparative groups 1 to 2 are all worse than those of the examples 2 and 12 to 14, and the effects on the above aspects of piglets of the examples 12 to 14 are obviously better than those of the example 2, which shows that the types of the cellulase or the glucose oxidase in the application are different, the effects on the piglets are obviously different after the cellulase or the glucose oxidase is matched with other raw materials in the application, and the effects of the cellulase and the glucose oxidase respectively used in the examples 12 and 13 are the best.
In addition, the effects of the comparative groups 3-4 on the growth performance, the immune function and the diarrhea rate of piglets are all obviously worse than those of the example 2, and in addition, the comparison between the example 2 and the examples 12-14 shows that the cellulase and the glucose oxidase have certain synergistic effect in the application.
Test example 3
Taking the example 2 and the examples 14-16 as examples, the 4 groups of the composite plant extract feed additives are respectively added into the feed of piglets according to the same amount and fed to the piglets according to the method of the test example 1. The results show that the piglets in examples 14-16 have higher average daily gain and average daily feed intake, lower feed-to-weight ratio, better serum immunity index and lower diarrhea rate than the piglets in example 2. The cysteine is added into the enzymolysis system in the enzymolysis process, which is favorable for improving the immunocompetence of the piglets and promoting the growth of the piglets.
In conclusion, the composite plant extract feed additive provided by the preferred embodiment of the invention has the advantages of reasonable formula, natural components, no toxic or side effect and no drug resistance, is beneficial to regulating the organism immunity of live pigs, promotes the growth of the live pigs and reduces the morbidity. The preparation method is simple to operate, is rapid and efficient, and can enable the feed additive to have a good effect. The product can be used for preparing pig feed, and can significantly improve feed intake and immunity of pig, and reduce diarrhea.
The embodiments described above are some, but not all embodiments of the invention. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Claims (10)

1. The composite plant extract feed additive is characterized in that raw materials of the composite plant extract feed additive comprise 11-12 parts by weight of purslane, 6-8 parts by weight of stevia rebaudiana leaves, 3-5 parts by weight of acanthopanax, 1-3 parts by weight of astragalus membranaceus, 1-2 parts by weight of Chinese violet and 4-6 parts by weight of callicarpa bodinieri;
the plant extract feed additive is prepared by the following method: mixing the purslane, the stevia rebaudiana leaves, the acanthopanax, the astragalus membranaceus, the Chinese violet and the callicarpa bodinieri according to a ratio to obtain a primary material, mixing the primary material with water and a compound enzyme, performing enzymolysis, concentrating and drying;
the feed-liquid ratio of the primary material to the water is 1 g: 5-10mL and/or the weight ratio of the primary material to the complex enzyme is 100: 0.1-0.5;
the complex enzyme comprises cellulase and glucose oxidase; the weight ratio of the cellulase to the glucose oxidase is 1: 0.5-0.8;
the enzymolysis is carried out at 45-65 deg.C for 8-12 h.
2. The composite plant extract feed additive as claimed in claim 1, wherein the raw materials comprise 11.2-11.8 parts by weight of the purslane, 6.5-7.5 parts by weight of the stevia rebaudiana leaves, 3.5-4.5 parts by weight of the acanthopanax senticosus, 1.5-2.5 parts by weight of the astragalus membranaceus, 1.2-1.8 parts by weight of the Chinese violet and 4.5-5.5 parts by weight of the callicarpa bodinieri.
3. The composite plant extract feed additive as claimed in claim 2, wherein the raw materials comprise 11.5 parts by weight of the purslane, 7 parts by weight of the stevia rebaudiana leaves, 4 parts by weight of the acanthopanax senticosus, 2 parts by weight of the astragalus membranaceus, 1.5 parts by weight of the Chinese violet and 5 parts by weight of the callicarpa bodinieri.
4. A method for preparing a composite plant extract feed additive as claimed in any one of claims 1 to 3, comprising the steps of: mixing the purslane, the stevia rebaudiana leaves, the acanthopanax, the astragalus membranaceus, the Chinese violet and the callicarpa bodinieri according to a ratio to obtain a primary material, mixing the primary material with water and the complex enzyme, performing enzymolysis, concentrating and drying.
5. The preparation method according to claim 4, wherein cysteine is added into an enzymolysis system in the enzymolysis process, and the weight ratio of the cysteine to the cellulase is 0.2-0.4: 1.
6. the method of claim 4, wherein the concentration and drying are carried out by concentrating the enzymatic hydrolysate at 60-80 deg.C, and freeze-drying at-55 deg.C for 12-24 h.
7. The preparation method of claim 4, wherein the step of shearing, drying and crushing the primary material in sequence before mixing with water and the complex enzyme is further carried out, and then the primary material is sieved by a 30-50 mesh sieve.
8. The use of a composite plant extract feed additive as claimed in any one of claims 1 to 3, wherein the composite plant extract feed additive is used to prepare a pig feed.
9. The use according to claim 8, wherein the composite plant extract feed additive is added to the live pig feed in a proportion of 0.01-0.1 wt%.
10. The use according to claim 9, wherein the composite plant extract feed additive is added to the live pig feed in a proportion of 0.05 wt%.
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