CN110604757A - Compound preparation for strengthening body resistance and detoxifying, and preparation method and application thereof - Google Patents
Compound preparation for strengthening body resistance and detoxifying, and preparation method and application thereof 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)
- A61K36/19—Acanthaceae (Acanthus family)
- A61K36/195—Strobilanthes
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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)
- A61K36/29—Berberidaceae (Barberry family), e.g. barberry, cohosh or mayapple
- A61K36/296—Epimedium
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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)
- A61K36/31—Brassicaceae or Cruciferae (Mustard family), e.g. broccoli, cabbage or kohlrabi
- A61K36/315—Isatis, e.g. Dyer's woad
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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)
- A61K36/34—Campanulaceae (Bellflower family)
- A61K36/344—Codonopsis
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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)
- A61K36/46—Eucommiaceae (Eucommia family), e.g. hardy rubber tree
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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)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/481—Astragalus (milkvetch)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/37—Extraction at elevated pressure or temperature, e.g. pressurized solvent extraction [PSE], supercritical carbon dioxide extraction or subcritical water extraction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
Abstract
The invention discloses a compound preparation for strengthening body resistance and detoxifying, a preparation method and application thereof, wherein the preparation method comprises the following steps: s1: respectively carrying out amplification culture on lactobacillus and bacillus and aspergillus niger, and carrying out volume ratio on the amplified bacterium liquid of the lactobacillus bacterium liquid: and (3) bacillus liquid: the aspergillus niger bacterial liquid is 1-4: 2-4: 1-3, mixing to obtain a compound bacterial liquid; s2: mixing 2-6 parts of astragalus, 2-6 parts of isatis root and 2-4 parts of epimedium according to parts by weight, and sterilizing at the temperature of 110-; s3: mixing the compound bacterial liquid and the sterilized Chinese herbal medicine mixture according to the weight ratio of liquid to solid of 15-30: 1, mixing and then fermenting in a closed manner for 72-96 h; s4: continuously mixing 0.1-1.0% of cellulolytic enzyme and 0.1-1.0% of lysostaphin into the fermented mixture, and performing CO treatment for 4-8 hr2The effective components of the finished product prepared by supercritical extraction and freeze drying of the extract are improved, and the organism immunity of the livestock is improved.
Description
Technical Field
The invention relates to the technical field of large-scale breeding of livestock, in particular to a compound preparation for strengthening body resistance and detoxifying, and a preparation method and application thereof.
Background
In the process of livestock and poultry and aquaculture, the immune function is easily inhibited due to long-term extensive stress factors. Taking a pig as an example, in this case, various pathogenic factors such as viruses, bacteria and protozoa can invade and destroy the physiology and immune system functions of the pig, free radicals generated by organ and tissue metabolism and acidic substances such as lactic acid, keto acid and uric acid can cause long-term acid bias of pig body fluid (pH is between 7.31 and 7.36), and in this case, the blood viscosity of the pig is high, the fluidity is low, and the effect of cells can be weakened; the oxygen carrying and nutrient carrying capacity of red blood cells is reduced, the enzymatic reaction efficiency is reduced, the metabolism is slowed down, the physiological function is declined, the immune system function is low, and particularly, the nutrition cannot be well absorbed. Various chronic diseases which can not be cured for a long time can be caused.
Moreover, with the development of the breeding industry in China, extensive family feeding is gradually changed into specialized, intensive and high-density feeding, and the pig raising level is obviously improved. During the growth process of the pigs, the chronic poisoning, the increase of drug resistance and the reduction of resistance are caused by continuously and excessively adding antibiotics for a long time. In each breeding link, such as frequent herd combining and herd transferring, frequent material changing and people changing, and the like, the breeding environment has dirty air, drinking water pollution, overproof pathogenic microorganisms, and the like, and the pathogens and toxins firstly damage the digestive tract of animals in the animal body to cause digestive tract inflammation or ulcer, which is manifested as digestive disorders such as vomit, constipation, diarrhea and the like, and secondly damage the integrity of the respiratory tract mucous membrane to destroy the immune barrier of the respiratory tract mucous membrane, which is manifested as respiratory tract syndrome.
Furthermore, in the current breeding mode, the normal immune response of pigs is affected by long-term multiple-vaccine-disorganization, so that immune interference, immune tolerance and immune failure are caused. Heavy metal and mycotoxin in the feed exceed standards, and reproductive hormone is abused for a long time, so that the physical ability of the sow is reduced, the resistance is low, and the uterine contraction is weak; flushing and swelling vulva, vomiting, anorexia and diarrhea of the sow; the multiparous sow has no estrus, false estrus, abortion and stillbirth, insufficient milk, overlong birth process or repeated mating infertility. In obsolete sows which are frequently infested, more than 80 percent of the obsolete sows are caused by the reasons, so that the problems of the reduction of the immunity of animal organisms, the reduction of the quality of livestock and poultry products, the enhancement of the drug resistance of bacteria and the like are solved.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides the compound preparation for strengthening body resistance and detoxifying, which improves the effective components of the compound preparation for strengthening body resistance and detoxifying and improves the organism immunity of livestock.
A preparation method of a compound preparation for strengthening body resistance and detoxifying comprises the following steps:
s1: respectively carrying out amplification culture on lactobacillus and bacillus and aspergillus niger, and then, carrying out amplification culture on the expanded bacterium liquid according to the volume ratio of the lactobacillus bacterium liquid: and (3) bacillus liquid: the ratio of the aspergillus niger liquid is 1-4: 2-4: 1-3, and preparing a compound bacterial liquid;
s2: mixing 2-6 parts of astragalus, 2-6 parts of isatis root and 2-4 parts of epimedium according to parts by weight, and sterilizing at the temperature of 110-;
s3: mixing the compound bacterial liquid and the sterilized Chinese herbal medicine mixture according to the weight ratio of liquid to solid of 15-30: 1, uniformly mixing, and fermenting in a sealed manner for 72-96 h;
s4: continuously mixing the fermentation mixture with 0.1-1.0% of cellulolytic enzyme and 0.1-1.0% of lysostaphin, and performing CO treatment for 4-8 hr2Performing supercritical extraction, and freeze drying the extract to obtain the finished product.
By adopting the scheme, the Chinese herbal medicine is sterilized at high temperature before fermentation, so that the influence of mixed bacteria on the fermentation process is avoided. During fermentation, lactic acid, bacillus and aspergillus niger are selected as fermentation strains, almost complete sets of decomposition enzymes aiming at the botanical raw materials are contained, the decomposition enzymes are microorganisms with the strongest decomposition capability in the natural world, the toxicity of the Chinese herbal medicines is reduced through the powerful biotransformation function of the microorganisms, and natural plant cells are subjected to enzymolysis, so that the effective components in the Chinese herbal medicines are fully released freely and enriched. After fermentation, the added cellulolytic enzyme and lysostaphin can selectively degrade natural macromolecules, plant polypeptides and the like in the Chinese herbal medicines into micromolecular substances which are easily absorbed and utilized by organisms, such as various glycosides, alkaloids, polysaccharides, organic sugars, volatile oil and other immune active substances. CO 22The supercritical extraction process has low extraction temperature, and can reduce the damage to effective components of Chinese medicinal materials as much as possible. The obtained product contains multiple immunocompetent substances, and can regulate nerve, body fluid and cell molecules of organism in all directions, and activate cell and humoral immunity system of organismHas effects in improving disease resistance of livestock and fowl, clearing away heat and toxic materials, strengthening body resistance, and eliminating pathogenic factors.
Preferably, the expanded culture of aspergillus niger in step S1 is: aspergillus niger is subjected to amplification culture with culture solution composed of 8-12% by mass of MAIZHU extract and 5-10% by mass of defatted soybean powder, pH6.5-6.8, and continuously fermented for 70-75 h.
By adopting the scheme, the special culture solution is prepared for the aspergillus niger, so that the culture effect of the aspergillus niger is improved.
Preferably, the Chinese herbal medicines in the step S2 further include, by weight, 2-4 parts of eucommia ulmoides and 2-4 parts of codonopsis pilosula.
By adopting the scheme, the eucommia ulmoides is sweet in taste and warm in nature, has the effects of tonifying liver and kidney, strengthening tendons and bones, regulating Chong and ren meridians, strengthening channels and preventing miscarriage, and has the effects of tonifying middle-jiao and Qi, invigorating spleen and benefiting lung. Radix Codonopsis has effects of enhancing immunity, dilating blood vessel, lowering blood pressure, improving microcirculation, and improving hemopoiesis function. The addition of eucommia bark and radix codonopsitis can further improve the types of effective components in the compound preparation, and further can further improve the drug effect.
Preferably, the optimal temperature for the sealed fermentation in step S3 is 32-35 deg.C, and the enzyme production temperature is 28-30 deg.C.
Preferably, a synergist is added during the closed fermentation in step S3, wherein the synergist is a mineral element Cu2+、Co2+、Mg2+、Fe2+、Zn2+、Ca2+The water-soluble compound of (a) is an aqueous solution prepared by the following element substances in ratio: cu2+：Co2+：Mg2+：Fe2+：Zn2+：Ca2+＝1.0-6.25：1.5-5.0：20-100：18-89：38.5-115：250-500。
Preferably, the synergist is made of CaCl2，ZnSO4，Fe SO4，CuSO4，Co(NO3)2，MgSO4Preparing the aqueous solution.
By adopting the scheme, the mineral elements can provide various nutrient components for the growth of microorganisms, improve the quantity of viable bacteria in the fermentation liquor, improve the stability and activity of the performance of the composite preparation, shorten the fermentation period and reduce the fermentation cost.
Preferably, the prepared finished product dosage form is one of granules, effervescent granules, dispersible tablets, effervescent tablets, film-coated tablets, pills, capsules, soft capsules, dripping pills, oral liquid and injection.
Object two of the present invention: provides a compound preparation for strengthening body resistance and detoxifying, which is prepared by the preparation method.
The third purpose of the invention is that: provides the application of the compound preparation for strengthening body resistance and detoxifying, and is applied to the field of livestock breeding.
Preferably, the feed additive is used for improving the immunity of pigs.
In conclusion, the invention has the following beneficial effects:
1. in the fermentation process, lactic acid bacteria, bacillus and aspergillus niger are selected, the synergistic effect of Chinese herbal medicines and probiotics is utilized, namely the Chinese herbal medicine active ingredients promote the proliferation efficiency of the probiotics, the probiotics carry out biotransformation on the Chinese herbal medicines to generate probiotic mycoprotein antibodies, oligosaccharides and derivatives thereof, and the probiotic mycoprotein antibodies, the oligosaccharides and the derivatives thereof have the effects of resisting inflammation and infection and promoting the propagation of intestinal beneficial flora and also have the effect of activating the immune function of the organism. Preventing and treating various viral respiratory diseases, hyperpyrexia syndrome and the like of pigs. The drug effect of active ingredients in the Chinese herbal medicine is improved;
2. the added cellulolytic enzyme and lysostaphin can selectively decompose natural macromolecules, plant polypeptides and the like in the Chinese herbal medicine to generate a plurality of immunocompetent substances such as glycosides, alkaloids, polysaccharides, organic sugars, volatile oil and the like, and the immunocompetent substances are matched with decomposition fermentation products to further improve the types and the activities of effective components in the Chinese herbal medicine and improve the utilization rate of the Chinese herbal medicine;
3. selecting CO2The supercritical extraction process has low extraction temperature, and can reduce the damage to the effective components of the Chinese herbal medicines as much as possible compared with the traditional Chinese herbal medicine decoction extraction mode;
4. the compound preparation of the invention is not added with preservative, and is a novel compound Chinese herbal medicine microecological preparation which is efficient, green, residue-free, antibiotic-free and antibacterial. Can strongly improve immunity and construct multiple immunity barriers of livestock. Can be used as the first choice medicine for viral and bacterial respiratory diseases, and has effects of rapidly activating organism immunity, starting disease-resistant system, improving organism immunity and disease resistance, and reducing death rate. Has wide action range and quick curative effect, and can be added for a long time.
Drawings
FIG. 1 shows histological sections (400-fold) of livers of various groups of experimental piglets in a safety test for pigs as a target animal;
FIG. 2 is a histological section of spleen of each experimental group of piglets in safety test for target animal pigs (400-fold);
fig. 3 shows kidney histological sections (400-fold) of each group of experimental piglets in the safety test for the target animal pig.
Detailed Description
Example 1
A preparation method of a compound preparation for strengthening body resistance and detoxifying comprises the following steps:
s1: inoculating lactobacillus (200 hundred million CFU/ml) into MRS liquid culture solution for amplification culture, wherein the temperature of a gas bath shaking table is 200rmp, the fermentation is stopped when the pH of the bacterium solution reaches 5.0, and the fermentation time is 16 h; inoculating bacillus (1000 hundred million CFU/ml) to LB culture solution for amplification culture, and culturing for 24h at 35 ℃ in a gas bath shaker at 200rmp to ensure that the absorbance OD value of the bacterial solution reaches 4; carrying out amplification culture on Aspergillus niger by using a culture solution consisting of 8% by mass of a salt drum extract and 5% by mass of defatted soybean powder, carrying out continuous fermentation for 70h at a pH of 6.5, wherein the concentrated fermentation clear solution contains 3000U/ml of saccharifying enzyme and 7000U/ml of alpha-amylase; and (3) adding the expanded bacterial liquid into the lactic acid bacteria liquid according to the volume ratio: and (3) bacillus liquid: the ratio of the aspergillus niger liquid is 1: 4: 3, mixing to obtain a compound bacterial liquid;
s2: taking 2 parts of astragalus, 2 parts of isatis root and 4 parts of epimedium according to the parts by weight, uniformly stirring, and sterilizing at 110 ℃ for 20 min;
s3: mixing the compound bacterial liquid and the sterilized Chinese herbal medicine mixture according to the liquid-solid weight ratio of 15: 1, uniformly mixing, and fermenting in a closed manner for 72 hours at the optimum temperature of 32 ℃ and the enzyme production temperature of 28 ℃;
s4: continuously mixing the fermentation mixture with 0.1% of cellulolytic enzyme and the total weight of the mixtureLysostaphin accounting for 0.1 percent of the total mass of the mixture, and performing CO treatment after 4 hours2Performing supercritical extraction;
s5: and (2) putting the concentrated clear paste obtained by extraction into a vacuum freeze drying device, freeze drying at-10 ℃ to obtain dry paste with the water content of less than or equal to 6%, crushing the dry paste, sieving the dry paste by a sieve of 80 meshes to obtain dry paste powder, adding dextrin into the dry paste powder, uniformly mixing the dry paste powder in a groove-shaped mixer, adding corn starch slurry to prepare soft materials, wherein the mass fractions of dextrin and corn starch in the total mixture are respectively 30% and 20%, granulating the soft materials by a swing granulator to prepare wet granules, and drying, granulating and subpackaging the wet granules to obtain the granular preparation type strengthening and detoxifying composite preparation with the water content of less than or equal to 5%.
Example 2
A preparation method of a compound preparation for strengthening body resistance and detoxifying comprises the following steps:
s1 inoculating lactobacillus (200 hundred million CFU/ml) into MRS liquid culture solution for amplification culture, wherein the temperature of a gas bath shaker is 300rmp, the fermentation is stopped when the pH of the bacterium solution reaches 5.2, and the fermentation time is 20 h; inoculating bacillus (1000 hundred million CFU/ml) to LB culture solution for amplification culture, and culturing for 36h at 37 ℃ by using a gas bath shaker at 300rmp to ensure that the absorbance OD value of the bacterial solution reaches 5; carrying out amplification culture on Aspergillus niger by using a culture solution consisting of 10% by mass of a salt drum extract and 8% by mass of defatted soybean powder, carrying out continuous fermentation for 72h at a pH of 6.6, wherein the concentrated fermentation clear solution contains 3000U/ml of saccharifying enzyme and 7000U/ml of alpha-amylase; and (3) adding the expanded bacterial liquid into the lactic acid bacteria liquid according to the volume ratio: and (3) bacillus liquid: the ratio of the aspergillus niger liquid is 4: 2: 3, mixing to obtain a compound bacterial liquid;
s2: taking 4 parts of astragalus, 4 parts of isatis root and 3 parts of epimedium according to parts by weight, uniformly stirring, and sterilizing at the high temperature of 120 ℃ for 30 min;
s3: mixing the compound bacterial liquid and the sterilized Chinese herbal medicine mixture according to the weight ratio of liquid to solid of 20: 1, uniformly mixing, and fermenting in a closed manner for 82 hours at the optimum temperature of 34 ℃ and the enzyme production temperature of 29 ℃;
s4: continuously mixing the zymolysis mixture with the cellulose decomposition enzyme accounting for 0.4 percent of the total mass of the mixture and the lysostaphin accounting for 0.4 percent of the total mass of the mixture,after 6h, CO was carried out2Performing supercritical extraction;
s5: and (2) putting the concentrated clear paste obtained by extraction into a vacuum freeze drying device, freeze drying at-10 ℃ to obtain dry paste with the water content of less than or equal to 6%, crushing the dry paste, sieving the dry paste by a sieve of 80 meshes to obtain dry paste powder, adding dextrin into the dry paste powder, uniformly mixing the dry paste powder in a groove-shaped mixer, adding corn starch slurry to prepare soft materials, wherein the mass fractions of dextrin and corn starch in the total mixture are respectively 30% and 20%, granulating the soft materials by a swing granulator to prepare wet granules, and drying, granulating and subpackaging the wet granules to obtain the granular preparation type strengthening and detoxifying composite preparation with the water content of less than or equal to 5%.
Example 3
A preparation method of a compound preparation for strengthening body resistance and detoxifying comprises the following steps:
s1: inoculating lactobacillus (200 hundred million CFU/ml) into MRS liquid culture solution for amplification culture, wherein the temperature of a gas bath shaking table is 350rmp, the fermentation is stopped when the pH of the bacterium solution reaches 5.5, and the fermentation time is 24 h; inoculating bacillus (1000 hundred million CFU/ml) to LB culture solution for amplification culture, and culturing for 48h at 40 ℃ in a gas bath shaker at 350rmp to ensure that the absorbance OD value of the bacterial solution reaches 6; carrying out amplification culture on Aspergillus niger (100 hundred million CFU/ml) by using a culture solution consisting of 12% of wheat drum extract and 10% of defatted soybean powder, carrying out continuous fermentation for 75h by using a gas bath shaking table with the pH value of 6.8, and obtaining a concentrated fermentation clear solution containing 3000U/ml of saccharifying enzyme and 7000U/ml of alpha-amylase; and (3) adding the expanded bacterial liquid into the lactic acid bacteria liquid according to the volume ratio: and (3) bacillus liquid: the ratio of the aspergillus niger liquid is 4: 4: 1, mixing to obtain a compound bacterial liquid;
s2: taking 6 parts of astragalus, 6 parts of isatis root and 2 parts of epimedium according to parts by weight, uniformly stirring, and sterilizing at the high temperature of 130 ℃ for 40 min;
s3: mixing the compound bacterial liquid and the sterilized Chinese herbal medicine mixture according to the liquid-solid weight ratio of 30: 1, uniformly mixing, and fermenting in a closed manner for 96 hours at the optimum temperature of 35 ℃ and the enzyme production temperature of 30 ℃;
s4: continuously mixing the fermentation mixture with 1.0% of cellulolytic enzyme and 1.0% of lysostaphin, and performing CO treatment after 8 hr2Performing supercritical extraction;
s5: and (2) putting the concentrated clear paste obtained by extraction into a vacuum freeze drying device, freeze drying at-10 ℃ to obtain dry paste with the water content of less than or equal to 6%, crushing the dry paste, sieving the dry paste by a sieve of 80 meshes to obtain dry paste powder, adding dextrin into the dry paste powder, uniformly mixing the dry paste powder in a groove-shaped mixer, adding corn starch slurry to prepare soft materials, wherein the mass fractions of dextrin and corn starch in the total mixture are respectively 30% and 20%, granulating the soft materials by a swing granulator to prepare wet granules, and drying, granulating and subpackaging the wet granules to obtain the granular preparation type strengthening and detoxifying composite preparation with the water content of less than or equal to 5%.
Example 4
This example was carried out as in example 2, with the following differences: the Chinese herbal medicines in the step S2 further comprise 2 parts of eucommia ulmoides and 2 parts of codonopsis pilosula according to parts by weight.
Example 5
This example was carried out as in example 2, with the following differences: the Chinese herbal medicines in the step S2 further comprise 3 parts of eucommia ulmoides and 3 parts of codonopsis pilosula according to parts by weight.
Example 6
This example was carried out as in example 2, with the following differences: the Chinese herbal medicines in the step S2 further comprise 4 parts of eucommia ulmoides and 4 parts of codonopsis pilosula according to parts by weight.
Example 7
This example was carried out as in example 2, with the following differences: in the step S3, a synergist is added during closed fermentation, wherein the synergist is composed of a mineral element Cu2+、Co2+、Mg2+、Fe2+、Zn2+、Ca2+The water-soluble compound of (a) is an aqueous solution prepared by the following element substances in ratio: cu2+：Co2+：Mg2+：Fe2+：Zn2+：Ca2+1.0: 1.5: 20: 18: 38.5: 250, selecting water-soluble compound as CaCl2 2-4g/L，ZnSO4 0.5-1.5g/L，FeSO4 0.2-1.0g/L，CuSO4 0.01-0.08g/L，Co(NO3)2 0.02-0.06g/L，MgSO4 0.1-0.5g/L。
Example 8
This example was carried out as in example 2, with the following differences: in the step S5, a synergist is added during closed fermentation, wherein the synergist is composed of a mineral element Cu2+、Co2+、Mg2+、Fe2+、Zn2+、Ca2+The water-soluble compound of (a) is an aqueous solution prepared by the following element substances in ratio: cu2+：Co2+：Mg2+：Fe2+：Zn2+：Ca2+3.0: 3.5: 60: 55: 68: 375 and the water-soluble compound is selected from CaCl2 2-4g/L，ZnSO4 0.5-1.5g/L，FeSO4 0.2-1.0g/L，CuSO4 0.01-0.08g/L，Co(NO3)20.02-0.06g/L，MgSO4 0.1-0.5g/L。
Example 9
This example was carried out as in example 2, with the following differences: in the step S5, a synergist is added during closed fermentation, wherein the synergist is composed of a mineral element Cu2+、Co2+、Mg2+、Fe2+、Zn2+、Ca2+The water-soluble compound of (a) is an aqueous solution prepared by the following element substances in ratio: cu2+：Co2+：Mg2+：Fe2+：Zn2+：Ca2+6.25: 5.0: 100: 89: 115: 500, selecting water-soluble compound as CaCl2 2-4g/L，ZnSO4 0.5-1.5g/L，Fe SO4 0.2-1.0g/L，CuSO4 0.01-0.08g/L，Co(NO3)2 0.02-0.06g/L，MgSO4 0.1-0.5g/L。
Active ingredient detection
The data for the assay of the major ingredients of the finished products prepared in examples 1-9 are shown in Table 1.
Table 1 results of effective ingredient detection data.
As can be seen from examples 1-3 in Table 1, the preparation process has a significant effect on the amount of active ingredients in the finished product, and the preparation parameters in example 2 are superior; the icariin and astragaloside IV of the finished products prepared in examples 4 to 6 are similar to those of example 2, and the finished products prepared in examples 4 to 6 further contain various trace effective components such as sinigrin, adenosine, aucubin, codonopsis pilosula saponin, various amino acids and the like; the icariin and astragaloside IV contents of the finished products prepared in examples 7-9 are higher than those of example 2, because the addition of the synergist provides various nutrient components for the growth of microorganisms, improves the number of viable bacteria in fermentation liquor, and further improves the fermentation decomposition effect.
Safety test for target animal pig
The sample prepared in example 2 was selected for safety tests to confirm the safety of the composite formulation prepared by the preparation method of the present invention, as follows.
Purpose of the test
According to the requirements of the technical Specification for experimental clinical tests (trial) of the Ministry of agriculture and the technical guide principle of clinical tests of traditional Chinese medicines and natural medicines for veterinary use of the Ministry of agriculture, the safety and the safe dose of the samples on the clinical application of the target animal pigs are evaluated.
2 materials and methods
2.1 materials
Test drugs: example 2 sample prepared.
Experimental animals: 40 weaned piglets of 50 days old are of unlimited varieties and are half female and half male.
2.2 Experimental grouping and feeding
Piglets were kept and observed in a standard animal house (certified by the institutional animal care committee in beijing) for 2 days after the purchase of piglets, and 40 piglets were randomly divided into 4 groups of 10 piglets each. 3 dose groups of the drug were set according to 1, 3, 5 times (i.e., 0.4g/kg, 1.2g/kg, 2.0g/kg) of the maximum pharmacodynamic dose, while a blank control group without drug administration was set. During the test period, the feed and drinking water were fed according to the normal feeding schedule, and the temperature and humidity were maintained at 20 + -25 deg.C and 40-60% respectively. The administration mode adopts mixed feeding administration, 1 time per day and 15 days of continuous administration.
2.3 Effect evaluation index and method
The physical signs of the experimental piglets are as follows: the piglets were observed and recorded for feeding, mental and fecal (urine, feces) condition, and compared with the control group for sign difference.
Relative rate of weight gain: the piglets are weighed 1 time respectively before the beginning and after the end of the test, and the relative weight gain rate is calculated, wherein the relative weight gain rate is equal to the relative weight gain of a drug treatment group/the relative weight gain of a blank control group multiplied by l 00%.
Blood routine and blood biochemical indices: after the test is finished, each group of piglets draws blood to carry out conventional blood examination and biochemical blood index examination.
And (3) pathological observation: after the experiment is finished, the piglets are sacrificed, the liver, the spleen and the kidney are taken out for each group of dissection, the condition of each organ is observed, meanwhile, the tissue and the organ of the piglets are taken and weighed, and the organ coefficient is calculated, wherein the organ coefficient is the weight of the organ/the total weight multiplied by l 00%.
Pathological tissue section: corresponding lesion tissues are taken according to the lesion conditions of the piglets, and 3 piglets in each group are taken
Organ tissues are fixed by 10% formaldehyde solution, pathological sections are prepared, and the histopathological conditions are observed.
2.4 data analysis
Statistical software SPASS 18.0 is used for data processing, independent sample t test is adopted, test data are represented by mean values +/-standard deviation, and the difference is significant when P is less than or equal to 0.05.
3 results of the test
3.1 Observation of body signs
The piglets in each group in the test period have good overall condition, normal ingestion and good mental status, and do not die; on the 7 th day of the test, 2 pigs in the control group showed cough symptoms, and recovered to normal after 3 days; the physical signs of the pigs in the drug groups with 1 time, 3 times and 5 times of doses are normal.
3.2 relative rate of weight gain
Compared with the blank control group, the relative weight gain rate of the 1-time, 3-time and 5-time dose groups is increased, but the statistically significant difference (p >0.05) is not generated (Table 2).
Table 2 relative weight gain of the piglets tested (n 10).
Note: the data in the same row without shoulder marks showed no significant difference (p >0.05), and data in the same row showed significant difference (p <0.05) compared to the control group.
3.3 blood routine test results
Compared with a blank control group, the difference between the detection indexes is not significant (p is greater than 0.05), and each detection index of each group is within a normal range (Table 3).
Table 3 piglet blood general indices (n-10) were tested.
Note: the data in the same row without shoulder marks showed no significant difference (p >0.05), and data in the same row showed significant difference (p <0.05) compared to the control group.
3.4 Biochemical index test results of blood
Compared with a blank control group, the content of glutamic-oxaloacetic transaminase (GOT), urea nitrogen (BUN) and Total Protein (TP) in the serum of the 1-time dose group is obviously lower than that of the blank control group; the contents of glutamic-oxaloacetic transaminase (GOT), glutamic-pyruvic transaminase (GPT) and urea nitrogen (BUN) in the serum of the 3-time dose group are obviously lower than those of the blank control group; the contents of glutamic-pyruvic transaminase (GPT) and urea nitrogen (BUN) in the serum of the 5-time dose group are obviously lower than those of the blank control group; each group of data, although significantly different from the blank group, was still within the reference range (table 4).
Table 4 biochemical indicators of blood of the piglets tested (n ═ 10).
Note: row data without shoulder marks indicated no significant difference (p >0.05) and row data indicated significant difference compared to control (p < 0.05).
3.5 organ coefficients
There was no significant difference in the organ coefficients compared to the blank control group (table 5).
Table 5 organ coefficients (n ═ 10) for the experimental piglets.
Note: the data in the same row without shoulder marks showed no significant difference (p >0.05), and data in the same row showed significant difference (p <0.05) compared to the control group.
3.6 histopathological examination
After the test is finished, all piglets are sacrificed, no obvious lesion is found by the autopsy, and then the histological detection is carried out.
3.6.1 liver
The structure of each group of liver cells is clear and complete, the shapes and the sizes are consistent, and the cytoplasm is rich; the nucleus is round, centered and even with double or multiple cores; the liver cable structure of each group is clear. Referring to fig. 1, a is a blank control group, B is a l-fold dose group, C is a 3-fold dose group, and D is a 5-fold dose group.
3.6.2 spleen
Spleen tissues of each group are clear in color, and red marrow and white marrow have obvious boundaries; the number of lymphocytes is large, the shape is consistent, and the coloring is uniform. Referring to fig. 1, a is a blank control group, B is a l-fold dose group, C is a 3-fold dose group, and D is a 5-fold dose group.
3.6.3 Kidney
Each group of renal tubules has clear shape, uniform staining, clear renal tubule epithelial cell boundary and complete cell nucleus; the glomerulus structure is complete. Referring to fig. 1, a is a blank control group, B is a l-fold dose group, C is a 3-fold dose group, and D is a 5-fold dose group.
4 conclusion of the test
The general condition, the observation of visceral organs and the examination of blood physiological and biochemical indexes of the piglets are carried out in the experiment, so that the sample prepared in the embodiment 2 has no harmful effect on the piglets and has better safety.
Immunity enhancement test of compound preparation on target animal pig
The samples prepared in representative examples 2, 5 and 8 were selected to be subjected to an immunopotentiation test to confirm the effect of the complex formulation prepared by the preparation method of the present invention in enhancing the immunity of animals.
Purpose of the test
The clinical efficacy of the samples prepared in examples 2, 5 and 8 on enhancing the pig organism immunity was evaluated according to the requirements of the Ministry of agriculture, the technical Specification for clinical laboratory tests (trial) and the Ministry of agriculture, the technical guidelines for clinical tests of veterinary Chinese medicine and natural medicine.
2 materials and methods
2.1 materials
Test animals: for piglets, the number of male and female is 65 respectively, 15 +/-20 kg, and the porcine reproductive and respiratory syndrome antibody is negative, and the antibody is purchased from Dabei agriculture breeding technology Limited liability company in Tangshan; cyclophosphamide was purchased from Hubei Kangbao Tai Fine chemical Co., Ltd, lot number KBT 20170501; porcine reproductive and respiratory syndrome attenuated vaccine was purchased from bleegger hagaham, germany, lot no: 1245110-05; samples prepared in examples 2, 5, 8; lingqi kunjin vitamin was purchased from astragalis health products limited, shandong, and the batch number: 20170501.
2.2 groups of test animals
Referring to table 6, 130 piglets were randomly divided into 13 groups of 5 females/males each. Group 1 is blank, groups 2-3 are model, and groups 4-13 are drug groups. The blank group was not injected with cyclophosphamide, administered, and vaccinated; model group 1 cyclophosphamide injection, but no administration, no vaccination; model group 2 was vaccinated with cyclophosphamide injection, but no drug: the drug group 1 is injected with cyclophosphamide, inoculated with vaccine and administered with contrast drug; drug group 2-4 injections of cyclophosphamide, vaccination, different doses (0.1-0.4g/kg-b.w.) of the samples prepared in example 2; drug groups 5-7 were injected with cyclophosphamide, vaccinated, and given different doses (0.1-0.4g/kg-b.w.) of the samples prepared in example 5; drug groups 8-10 were injected with cyclophosphamide, vaccinated and given different doses (0.1-0.4g/kg-b.w.) of the samples prepared in example 8. The molding administration procedure of the cyclophosphamide is that the cyclophosphamide injection is injected intramuscularly at the ratio of 75 mg/kg.b.w. on the 1 st to 4 th days of the experiment, and the same amount of normal saline is injected into the cyclophosphamide-free group; the drug administration program is that the drugs and the doses listed in the table 6 are mixed and fed for 1 time every day on the 1 st to 10 th days of the test; the procedure for vaccination was intramuscular injection of porcine reproductive and respiratory syndrome vaccine on day 11 of the experiment and the non-vaccinated group injected with an equivalent amount of normal saline. The test lasts for 50 days, the pigs are fed normally, and the condition of the pig herds is observed and recorded every day.
Table 6 test groupings.
2.3 index evaluation
2.3.1 body weight determination
Pigs were weighed for each group at 0, 10, 25, 50 days of the trial, respectively, and the differences in weight of pigs for each group were compared.
2.3.2 blood routine, blood Biochemical assay and antibody level assay
Collecting peripheral blood of each group of pigs on 0, 20 and 40 days after immunization, and dividing the peripheral blood into two parts, wherein one part is used for carrying out conventional blood detection by using a full-automatic blood cell analyzer, and the other part is used for detecting GOT, GTP, ALP, BUN, TP and ALB by using a full-automatic biochemical analyzer.
And collecting peripheral blood of each group of pigs on 0, 7, 15, 25 and 40 days after immunization, separating serum, and detecting the porcine reproductive and respiratory syndrome antibody level of each group by using a porcine reproductive and respiratory syndrome antibody detection kit.
2.4 data analysis
The SPASS 18.0 statistical software is used for data processing, independent sample t test is adopted, test data are all expressed by mean values +/-standard deviation, and the difference P <0.05 is obvious.
2.5 test results
2.5.1 body weight
The daily weight gain was lower for the remaining test groups at 10 and 25 days compared to the blank group, and the daily weight gain for 50 days was essentially the same for each test group, but was statistically not significantly poor. The daily weight gain of the first 10 days is obviously reduced compared with that of a blank group in a model group 1, a model group 2, a medicine group 1-3, a medicine group 5-6 and a medicine group 8-9; compared with the model group 2, the daily gain of the drug group 4, the drug group 7 and the drug group 10 was significantly increased, as shown in table 7.
Table 7 daily gain of the experimental piglets at different time periods (n-10).
Note: indicates significant difference compared to the blank group (P < 0.05); Δ indicates a significant difference compared to model group 2 (P < 0.05).
2.5.2 blood routine results
The blood routine detection is carried out on each group of piglets at 0, 20 and 40 days after immunization respectively, and the detection results are as follows:
day 0 after immunization, the level of White Blood Cells (WBC) was reduced and significantly different in each of the remaining test groups compared to the blank group, since the immunosuppressive effect of yellow phosphoramide was still affected; the Red Blood Cell (RBC) levels of model group 1, drug group 4, drug group 7, and drug group 10 were all significantly higher than the blank group. Compared with the model group 2, the Red Blood Cell (RBC) levels of the drug groups 2 to 3, 5 to 6 and 8 to 9 were significantly reduced, and the Red Blood Cell (RBC) levels of the drug group 1, 4, 7 and 10 were significantly increased, as shown in table 8.
Table 8 immunization day 0 standard index of piglet blood (n-10) was tested.
Note: indicates significant difference compared to blank group (P < 0.05); Δ indicates significant difference compared to model group 2 (P < 0.05).
On day 20 post-immunization, the White Blood Cell (WBC) levels were significantly elevated in model group 2, drug groups 1-10, compared to the blank group. Model group 2 showed significantly higher White Blood Cell (WBC) levels compared to model group 1. The drug groups 1, 4, 7, 10 showed significantly higher Platelet (PLT) than the model group 2, as shown in table 9.
Table 9 standard indices of piglet blood (n 10) tested on day 20 post immunization.
Note: indicates significant difference compared to blank group (P < 0.05); # indicates a significant difference compared to model group 2 (P < 0.05); Δ indicates significant difference compared to model group 2 (P < 0.05).
On day 40 after immunization, the White Blood Cell (WBC) levels were significantly increased in each group except the model group 1, and the Red Blood Cell (RBC) and platelet (RLT) levels were significantly decreased in the drug group 1, as compared to the blank group. Model group 2 showed significantly higher White Blood Cell (WBC) levels compared to model group 1. The White Blood Cell (WBC) levels were significantly elevated for drug group 1, drug groups 3-4, drug groups 6-7, and drug groups 9-10 compared to model group 2. See table 10.
Table 10 standard indices of piglet blood (n 10) were tested on day 40 after immunization.
Note: indicates significant difference compared to blank group (P < 0.05); # indicates a significant difference (<0.05) compared to model group 2; Δ indicates significant difference compared to model group 2 (P < 0.05).
2.5.3 blood Biochemical index detection results
The blood biochemical detection is carried out on each group of piglets at 0, 10, 20 and 40 days after immunization respectively, and the detection results are as follows:
on day 0 after immunization, compared with the blank group, the levels of Total Protein (TP) of glutamic-oxaloacetic transaminase (GOT), Albumin (ALB) of model group 2, drug group 4, drug group 5, drug group 7, drug group 8 and drug group 10, drug group 2, drug group 5 and drug group 8 were all significantly reduced; alkaline phosphatase (ALP) in the group consisting of the model group 1, the model group 2, the drug group 1, the drug group 4, the drug group 7, and the drug group 10, glutamic-pyruvic transaminase (GPT) in the group consisting of the drug group 3, the drug group 6, and the drug group 9, and Albumin (ALB) level in the group consisting of the model group 2 were significantly increased. Urea Nitrogen (BUN), Albumin (ALB) levels were significantly reduced for model group 2 compared to model group 1; total Protein (TP) levels were significantly elevated. Compared with the model group 2, the glutamic-oxaloacetic transaminase (GOT) and glutamic-pyruvic transaminase (GPT) of the drug group 3, the drug group 6 and the drug group 9, the glutamic-oxaloacetic transaminase (GOT) level of the drug group 4, the drug group 7 and the drug group 10 were significantly increased, and the Total Protein (TP) and Creatinine (CRE) level of the drug group 2, the drug group 5 and the drug group 8 were significantly decreased, as shown in table 11.
Table 11 blood biochemical markers (n 10) were tested on day 0 post immunization.
Note: indicates significant difference compared to blank group (P < 0.05); # indicates a significant difference compared to model group 1 (P < 0.05); Δ indicates significant difference compared to model group 2 (P < 0.05).
On day 20 after immunization, the glutamic-pyruvic transaminase (GPT) levels of model group 1, glutamic-oxalacetic transaminase (GOT) and glutamic-pyruvic transaminase (GPT) of model group 2, glutamic-pyruvic transaminase (GPT) of drug group 3, drug group 6 and drug group 9, and glutamic-pyruvic transaminase (GPT) levels were all significantly reduced compared to the blank group; alkaline phosphatase (ALP) in the model group 1, the model group 2, the drug group 4, the drug group 7, and the drug group 10, urea nitrogen (BUN) in the model group 1, the model group 2, the drug group 1, the drug group 2, the drug group 5, and the drug group 8, and Total Protein (TP) level in the model group 2 were significantly increased. The Albumin (ALB) was significantly reduced in model group 2 compared to model group 1. Compared with the model group 2, the Total Protein (TP) levels of the drug group 2, the drug group 5 and the drug group 8, the urea nitrogen (BUN) levels of the drug group 3, the drug group 6 and the drug group 9 and the Total Protein (TP) levels are obviously reduced; glutamic-oxaloacetic transaminase (GOT) of drug group 1, urea nitrogen (BUN) and glutamic-oxaloacetic transaminase (GOT) of drug group 2, drug group 5, and drug group 8, and glutamic-oxaloacetic transaminase (GOT) of drug group 4 were significantly increased, as shown in table 12.
Table 12 blood biochemical markers (n 10) were tested on day 20 post immunization.
Note: indicates significant difference compared to blank group (P < 0.05); # indicates a significant difference compared to model group 2 (P < 0.05); Δ indicates significant difference compared to model group 2 (P < 0.05).
At day 40 after immunization, the Albumin (ALB) levels in model group 2 and glutamic-oxaloacetic transaminase (GOT) levels in drug group 2, drug group 5, and drug group 8 were significantly increased. Compared with the model 1, the glutamic-oxaloacetic transaminase (GOT) of the model 2 is significantly reduced, and the urea nitrogen (BUN) is significantly increased. Compared with the model group 2, the glutamic-oxalacetic transaminase (GOT) of the drug group 1, the drug group 2, the drug group 5, and the drug group 8 was significantly increased, and the level of glutamic-pyruvic transaminase (GPT) of the drug group 1 was significantly decreased. See table 13.
Table 13 blood biochemical markers (n 10) were tested on day 40 post immunization.
Note: indicates significant difference compared to blank group (P < 0.05); # indicates a significant difference compared to model group 1 (P < 0.05); Δ indicates significant difference compared to model group 2 (P < 0.05).
2.5.4 antibody detection results
And (3) collecting peripheral blood of the pigs on 0 th, 7 th, 15 th, 25 th and 40 th days after immunization, separating serum, detecting the porcine reproductive and respiratory syndrome antibodies of each group by using a porcine reproductive and respiratory syndrome antibody detection kit, and judging the porcine reproductive and respiratory syndrome antibodies to be positive if the detection result of the kit antibody level is more than 0.4 and judging the porcine reproductive and respiratory syndrome antibodies to be negative if the detection result is less than 0.4. Antibody levels were analyzed as follows:
before immunization, there was no significant difference between the test groups compared to the blank group. 7 days after immunization: compared with the blank group, the antibody levels of the model 2 group, the drug group 1, the drug group 4, the drug group 7 and the drug group 10 are obviously increased; compared with the model group 2, the antibody levels of the drug group 4, the drug group 7 and the drug group 10 were significantly increased. 15 days after immunization: compared with the blank group, the antibody levels of the model group 2 and the drug groups 1-10 are obviously increased; model group 2 antibody levels were significantly higher than model group 1; compared with the model group 2, the antibody levels of the drug groups have no significant difference. 25 days after immunization: compared with a blank group, the antibody levels of the model group 2 and the drug groups 1-10 are obviously increased; the antibody levels of the model group 2, the model group 5 and the model group 8 are obviously higher than that of the model group 1; compared with the model group 2, the antibody levels of the drug groups 3-4, 6-7 and 9-10 are obviously increased. 40 days after immunization: compared with the blank group, the antibody levels of the model group 2 and the drug groups 1-10 are obviously increased; model group 2 antibody levels were significantly higher than model group 1; compared with the model group 2, the antibody levels of the drug group 4, the drug group 7 and the drug group 10 were significantly increased, as shown in table 14.
Table 14 antibody level results.
Note: indicates significant difference compared to blank group (P < 0.05); # indicates a significant difference compared to model group 1 (P < 0.05; Δ indicates a significant difference compared to model group 2 (P < 0.05).
2.6 conclusion
The analysis of the results shows that the compound preparation samples prepared in examples 2, 5 and 8 have the effect of improving the titer of the porcine reproductive and respiratory syndrome vaccine antibody, and the effect is better than that of a control medicament when the administration dosage is (0.2-0.4 g/kg-b.w.). The effect of the composite preparation samples prepared in examples 5 and 8 is better than that of example 2.
The above-mentioned embodiments are merely illustrative and not restrictive, and those skilled in the art can modify the embodiments without inventive contribution as required after reading this specification, but only fall within the scope of the claims of the present invention.
Claims (10)
1. A preparation method of a compound preparation for strengthening body resistance and detoxifying is characterized in that: the method comprises the following steps:
s1: respectively carrying out amplification culture on lactobacillus and bacillus and aspergillus niger, and then, carrying out amplification culture on the expanded bacterium liquid according to the volume ratio of the lactobacillus bacterium liquid: and (3) bacillus liquid: the ratio of the aspergillus niger liquid is 1-4: 2-4: 1-3, and preparing a compound bacterial liquid;
s2: mixing 2-6 parts of astragalus, 2-6 parts of isatis root and 2-4 parts of epimedium according to parts by weight, and sterilizing at the temperature of 110-;
s3: mixing the compound bacterial liquid and the sterilized Chinese herbal medicine mixture according to the weight ratio of liquid to solid of 15-30: 1, uniformly mixing, and fermenting in a sealed manner for 72-96 h;
s4: further mixing the amount of the mixture into the fermentation mixtureCellulolytic enzyme accounting for 0.1-1.0% of the total mass of the mixture and lysostaphin accounting for 0.1-1.0% of the total mass of the mixture, and performing CO treatment after 4-8h2Performing supercritical extraction, and freeze drying the extract to obtain the finished product.
2. The method for preparing a compound preparation for strengthening body resistance and removing toxic substance according to claim 1, wherein the expanded culture of aspergillus niger in step S1 is: aspergillus niger is subjected to amplification culture with culture solution composed of 8-12% by mass of MAIZHU extract and 5-10% by mass of defatted soybean powder, pH6.5-6.8, and continuously fermented for 70-75 h.
3. The method for preparing a compound preparation for strengthening body resistance and removing toxicity according to claim 1, wherein the method comprises the following steps: the Chinese herbal medicines in the step S2 further comprise 2-4 parts of eucommia ulmoides and 2-4 parts of codonopsis pilosula according to parts by weight.
4. The method for preparing a compound preparation for strengthening body resistance and removing toxicity according to claim 1, wherein the method comprises the following steps: in step S3, the optimum temperature for sealed fermentation is 32-35 deg.C, and the enzyme production temperature is 28-30 deg.C.
5. The method for preparing a compound preparation for strengthening body resistance and removing toxicity according to claim 1, wherein the method comprises the following steps: in the step S3, a synergist is added during closed fermentation, wherein the synergist is composed of a mineral element Cu2+、Co2+、Mg2+、Fe2+、Zn2+、Ca2+The water-soluble compound of (a) is an aqueous solution prepared by the following element substances in ratio: cu2+：Co2+：Mg2+：Fe2+：Zn2+：Ca2+=1.0-6.25：1.5-5.0：20-100：18-89：38.5-115：250-500。
6. The method for preparing a compound preparation for strengthening body resistance and removing toxicity according to claim 5, wherein the method comprises the following steps: the water-soluble compound is CaCl2，ZnSO4，FeSO4，CuSO4，Co(NO3)2，MgSO4。
7. The method for preparing a compound preparation for strengthening body resistance and removing toxicity according to claim 1, wherein the method comprises the following steps: the prepared finished product is one of granules, effervescent granules, dispersible tablets, effervescent tablets, film-coated tablets, pills, capsules, soft capsules, dripping pills, oral liquid and injection.
8. A complex formulation for strengthening body resistance and removing toxic substance prepared by the preparation method of any one of claims 1 to 7.
9. The use of the complex formulation for strengthening body resistance and removing toxic substance of claim 8, wherein: is applied to the field of livestock breeding.
10. The use of the complex formulation for strengthening body resistance and removing toxic substance as set forth in claim 9, wherein: can be used for improving pig immunity.
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