CN111802511A - Functional traditional Chinese medicine residue feed additive, preparation method and application - Google Patents
Functional traditional Chinese medicine residue feed additive, preparation method and application Download PDFInfo
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- CN111802511A CN111802511A CN202010709746.5A CN202010709746A CN111802511A CN 111802511 A CN111802511 A CN 111802511A CN 202010709746 A CN202010709746 A CN 202010709746A CN 111802511 A CN111802511 A CN 111802511A
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- artemisia annua
- feed additive
- folium artemisiae
- corn steep
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
-
- 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)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
- A61K36/282—Artemisia, e.g. wormwood or sagebrush
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/12—Antidiarrhoeals
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/14—Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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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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- Natural Medicines & Medicinal Plants (AREA)
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- Animal Behavior & Ethology (AREA)
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Abstract
The invention belongs to the technical field of cultivation, and discloses a functional traditional Chinese medicine feed additive, a preparation method and application thereof, wherein the feed additive comprises the following main materials: artemisia annua plant material, auxiliary materials and zymophyte; is prepared by mixing and fermenting artemisia annua substances with auxiliary materials and fermentation microbial inoculum; for addition to feed. According to the invention, the artemisia annua is fermented by the zymophyte to prepare the feed additive, and the feed added with the additive can effectively reduce the overfeeding condition of laying hens, improve the glossiness of eggshells, increase the brightness of the eggshells, obviously improve the red manure condition, form and thin the manure, and effectively relieve the enterovirus syndrome of livestock and poultry. The functional traditional Chinese medicine feed additive is suitable for being added into feed to feed livestock and poultry.
Description
Technical Field
The invention belongs to the technical field of breeding, and relates to a feed additive, in particular to a functional traditional Chinese medicine feed additive, a preparation method and application.
Background
Artemisia annua, also known as Artemisia annua, is a traditional Chinese medicine and belongs to an annual Compositae plant. In the traditional Chinese medicine treatment, the artemisia annua is often used for treating symptoms such as fever abatement, cold treatment, summer heat removal and the like, and particularly has the advantages of high efficiency, low toxicity and the like in the aspect of treating cerebral malaria. The medicinal value of the artemisia annua has a long history, and as early as 340 yuan, the artemisia annua extract is used for treating malaria, which is mentioned in elbow backup acute prescription written by pueraria flood of doctors in the east jin. In 1972, Chinese researchers separated and extracted an effective component for treating malaria from Artemisia annua for the first time, and named the effective component as artemisinin. The artemisinin has various pharmacological actions of resisting malaria, regulating immunity, resisting tumor, resisting fungi and the like. In view of the good anti-malaria treatment effect of artemisinin, the world health organization recommends the artemisinin as the first choice drug for resisting malaria.
Recent estimates of data from the world health organization indicate that there are 2.12 billion malaria patients in 2015, 42.9 million of whom die, about 1/2 worldwide with malaria risk, and 91 countries and regions with persistent malaria transmission. The artemisinin is taken as the first-line medicament for treating malaria which is the most safe and effective at present, and has remarkable medical and commercial values. The artemisia annua leaves are used as a main raw material for extracting the artemisinin, and the generated artemisia annua dregs and stems which cannot be utilized are thousands of tons. The herba Artemisiae Annuae contains effective components such as arteannuic acid, dihydroarteannuin, arteannuin, and arteannuin, in addition to antimalarial components such as arteannuin, arteannuin sodium, and artemether. The residual artemisia annua dregs and stems after extracting the artemisinin by pharmaceutical enterprises also contain a plurality of other effective components besides part of the artemisinin, the medicinal value of the effective components is not fully utilized, and if the effective components are directly discarded or burnt, the waste and the environmental pollution are caused. The artemisia annua dregs and stems can be made into feed, but because the artemisia annua dregs and stems contain a large amount of fibers and have poor palatability, inedibility or dyspepsia can be easily caused, so that the growth rate of the livestock is reduced, and the loss of farmers can be caused.
Disclosure of Invention
The invention aims to provide a functional traditional Chinese medicine feed additive for relieving enterovirus syndrome of livestock and poultry;
the invention also aims to provide a preparation method of the functional traditional Chinese medicine feed additive;
the invention also aims to provide an application of the functional traditional Chinese medicine feed additive.
In order to achieve the purpose, the invention adopts the technical scheme that:
a functional Chinese medicinal feed additive comprises effective components of herba Artemisiae Annuae, adjuvant and zymocyte agent; wherein the dosage of the raw materials adopts any one scheme or the combination of at least two schemes from a) to g),
a) the artemisia annua substances are 20-30 parts of artemisia annua dregs;
the auxiliary materials comprise: 3-8 parts of naked flower purple bead powder, 3-8 parts of dried folium artemisiae argyi/6-16 parts of fresh folium artemisiae argyi, 3-8 parts of dried corn steep liquor/6-16 parts of corn steep liquor, 8-12 parts of soybean meal, 2-4 parts of brown sugar and 1-3 parts of mountain flour;
the total amount of the zymophyte agent is more than or equal to 10 hundred million CFU/g;
the water content of the raw material is 30-50%;
wherein the artemisia annua dregs are obtained by extracting artemisinin from artemisia annua leaves in the industry and are commonly called artemisia annua leaves dregs; the dried folium Artemisiae Argyi is sun-dried folium Artemisiae Argyi, and the fresh folium Artemisiae Argyi is fresh folium Artemisiae Argyi;
b) the artemisia annua substance is 10-20 parts of artemisia annua dry leaves by weight
The auxiliary materials comprise: 3-8 parts of naked flower purple bead powder, 3-8 parts of dried folium artemisiae argyi/6-16 parts of fresh folium artemisiae argyi, 3-8 parts of dried corn steep liquor/6-16 parts of corn steep liquor, 8-12 parts of soybean meal, 2-4 parts of brown sugar and 1-3 parts of mountain flour;
the total amount of the zymophyte agent is more than or equal to 10 hundred million CFU/g;
the water content of the raw material is 30-50%;
c) the artemisia annua substances are 30-50 parts of artemisia annua dry whole plants in parts by weight;
the auxiliary materials comprise: 3-8 parts of naked flower purple bead powder, 3-8 parts of dried folium artemisiae argyi/6-16 parts of fresh folium artemisiae argyi, 3-8 parts of dried corn steep liquor/6-16 parts of corn steep liquor, 8-12 parts of soybean meal, 2-4 parts of brown sugar and 1-3 parts of mountain flour;
the total amount of the zymophyte agent is more than or equal to 10 hundred million CFU/g;
the water content of the raw material is 30-50%;
d) the artemisia annua substances are 50-70 parts of artemisia annua dry stems by weight;
the auxiliary materials comprise: 3-8 parts of naked flower purple bead powder, 3-8 parts of dried folium artemisiae argyi/6-16 parts of fresh folium artemisiae argyi, 3-8 parts of dried corn steep liquor/6-16 parts of corn steep liquor, 8-12 parts of soybean meal, 2-4 parts of brown sugar and 1-3 parts of mountain flour;
the total amount of the zymophyte agent is more than or equal to 10 hundred million CFU/g;
the water content of the raw material is 30-50%;
e) the artemisia annua substances are 40-60 parts of artemisia annua fresh leaves in parts by weight;
the auxiliary materials comprise: 3-8 parts of naked flower purple bead powder, 3-8 parts of dried folium artemisiae argyi/6-16 parts of fresh folium artemisiae argyi, 3-8 parts of dried corn steep liquor/6-16 parts of corn steep liquor, 8-12 parts of soybean meal, 2-4 parts of brown sugar and 1-3 parts of mountain flour;
the total amount of the zymophyte agent is more than or equal to 10 hundred million CFU/g;
the water content of the raw material is 30-50%;
f) the artemisia annua substances are 50-70 parts of fresh artemisia annua whole plants in parts by weight;
the auxiliary materials comprise: 3-8 parts of naked flower purple bead powder, 3-8 parts of dried folium artemisiae argyi/6-16 parts of fresh folium artemisiae argyi, 3-8 parts of dried corn steep liquor/6-16 parts of corn steep liquor, 8-12 parts of soybean meal, 2-4 parts of brown sugar and 1-3 parts of mountain flour;
the total amount of the zymophyte agent is more than or equal to 10 hundred million CFU/g;
the water content of the raw material is 30-50%;
g) 60-80 parts by weight of fresh stems of artemisia annua;
the auxiliary materials comprise: 3-8 parts of naked flower purple bead powder, 3-8 parts of dried folium artemisiae argyi/6-16 parts of fresh folium artemisiae argyi, 3-8 parts of dried corn steep liquor/6-16 parts of corn steep liquor, 8-12 parts of soybean meal, 2-4 parts of brown sugar and 1-3 parts of mountain flour;
the total amount of the zymophyte agent is more than or equal to 10 hundred million CFU/g;
the water content of the raw material was 40%.
Further, the fermentation inoculum is OD600And adding the fermentation broth of the corresponding zymophyte with the value of 1.2-1.8 to ensure that the water content in the raw material is 40%.
As a limitation, the fermentation inoculum includes at least one of lactobacillus plantarum, pichia pastoris, bacillus licheniformis, aspergillus niger, lactobacillus casei, trichoderma reesei, and bacillus subtilis.
The invention also provides a preparation method of the functional traditional Chinese medicine feed additive, which is obtained by mixing and fermenting the artemisia annua with auxiliary materials and fermentation bacteria.
As a limitation, the screening granularity of the artemisia annua dregs, the dried artemisia annua leaves, the whole dried artemisia annua herbs, the dried artemisia annua stems and stems, the callicarpa nudiflora powder, the dried artemisia argyi leaves, the dried corn steep liquor powder, the bean pulp, the brown sugar and the stone powder is more than or equal to 40 meshes;
wherein, the dry leaves, the dry grass and the dry stems and stems of the artemisia annua are respectively dried, crushed and sieved artemisia annua leaves, artemisia annua whole plants and artemisia annua stems and stems;
the fresh leaves, fresh whole plants, fresh stems and stems of Artemisia annua and fresh leaves of Artemisia argyi are respectively picked, cleaned and cut leaves, whole plants, stems and stems of Artemisia annua and Artemisia argyi.
As another limitation, the fermentation temperature is room temperature, and the fermentation time is 12-30 days.
As a third limitation, when the fermentation inoculum is a mixed inoculum, the fermentation inoculum is inoculated in a culture medium and cultured to OD600After the value is 1.2-1.8, mixing fermentation liquor corresponding to the obtained fermentation strain to obtain the fermentation strain;
when the zymophyte agent is a single agent, a zymophyte is taken and inoculated in a culture medium to be cultured to OD600Obtaining the product after the value is 1.2-1.8;
wherein, the fermentation inoculum with the best fermentation effect is a mixed inoculum, in particular to the following four: OD600When the value is 1.2-1.8, taking the volume ratio of 1: 1: 1, mixing a mixed microbial inoculum prepared by mixing lactobacillus plantarum fermentation liquor, saccharomycete fermentation liquor and bacillus licheniformis fermentation liquor, wherein the volume ratio of the mixed microbial inoculum is 1: 1: 1, mixing Aspergillus niger fermentation liquor, Lactobacillus casei fermentation liquor and Trichoderma reesei fermentation liquor to obtain a mixed microbial inoculum, wherein the volume ratio of the Aspergillus niger fermentation liquor to the Lactobacillus casei fermentation liquor to the Trichoderma reesei fermentation liquor is 1: 1: 1, mixing lactobacillus casei fermentation liquor, trichoderma reesei fermentation liquor and bacillus subtilis fermentation liquor to obtain a mixed microbial inoculum, and taking the mixture in a volume ratio of 1: 1: 1: 1 mixed microbial inoculum prepared by mixing lactobacillus plantarum fermentation liquor, bacillus licheniformis fermentation liquor, aspergillus niger fermentation liquor and bacillus subtilis fermentation liquor.
The invention also provides an application of the functional traditional Chinese medicine feed additive, and the functional traditional Chinese medicine feed additive is used for being added into feed for feeding livestock and poultry.
As a limitation, the feed added with the functional traditional Chinese medicine feed additive is used for relieving enterovirus syndrome of livestock and poultry.
Due to the adoption of the technical scheme, compared with the prior art, the invention has the technical progress that:
according to the invention, the artemisia annua substances are fermented by using the fermentation microbial inoculum, and the obtained functional traditional Chinese medicine feed additive contains crude proteins, fats and other substances which can be utilized by livestock and poultry, and also contains functional components in the artemisia annua such as dihydroartemisinic acid (A) and artemisinin B, so that the enterovirus syndrome of the livestock and poultry can be effectively relieved. By feeding the feed added with the functional traditional Chinese medicine feed additive, the over-eating condition of the laying hens is effectively reduced, the glossiness of the eggshells is improved, the brightness of the eggshells is increased, the condition of red manure is obviously improved, and the manure is formed and becomes fine. According to the invention, the artemisia annua substances are fermented by using the fermentation microbial inoculum, and relatively, the microbial inoculum can decompose cellulose in the artemisia annua substances into nutrients which can be absorbed by livestock and poultry, so that the artemisia annua substances are fermented by using the fermentation microbial inoculum, the cellulose content is reduced by 2.0-5.2%, the crude protein content is increased by 5-16%, the fat content is increased by more than 0.2%, four functional components including dihydroartemisinic acid, artemisinic acid, artemisinin and artemisinin in artemisia annua are all increased after fermentation, the feed intake of the livestock and poultry is increased by 25-30%, the quality of egg and meat is improved, and the palatability of the artemisia annua substances is improved. In particular, Artemisia annua dregs (commonly called leaves and dregs after artemisinin extraction from Artemisia annua leaves) and Artemisia annua stems are used as raw materials for fermentation, so that resource waste and environmental pollution are reduced, meanwhile, the problem that the Artemisia annua dregs are easy to cause indigestion of livestock and poultry is solved, enterovirus syndromes of the livestock and poultry are relieved, and the application range of the Artemisia annua dregs and stems is expanded.
The functional traditional Chinese medicine feed additive is suitable for being added into feed, and is further used for feeding livestock and poultry, especially livestock and poultry suffering from enterovirus syndrome.
Drawings
FIG. 1 is a graph of a complexing protein standard determined in example 58 of the present invention;
fig. 2 is a GC-MS analysis diagram of the functional Chinese herbal feed additive N10 measured in example 58 of the present invention, wherein a is a general diagram, 1 is dihydroartemisinic acid, 2 is artemisinic acid, 3 is artemisinin, 4 is artemisinin, B is an analysis diagram of dihydroartemisinic acid, C is an analysis diagram of artemisinic acid, D is an analysis diagram of artemisinin, and E is an analysis diagram of artemisinin;
FIG. 3 shows control group A and control group B in example 59 of the present invention1Group stool photo contrast;
FIG. 4 shows control group A and control group B in example 59 of the present invention1Photo comparison of eggs laid by the group.
Detailed Description
The present invention is further illustrated by the following specific examples, which are to be construed as merely illustrative, and not limitative of the remainder of the disclosure.
Lactobacillus plantarum (CGMCC11160), Pichia pastoris (CGMCC25688), Bacillus licheniformis (ATCC27811), Aspergillus niger (ATCC30785), Lactobacillus casei (CGMCC1580), Trichoderma reesei (ATCC56765) and Bacillus subtilis (CGMCC17740) are purchased from China center for culture collection or American type culture collection and stock ATCC (American type culture Collection).
Example 1A method for preparing a fermentation inoculum
11) Preparing glucose and water into 20% glucose aqueous solution, sterilizing at 121 deg.C under 0.101MPa for 20min to obtain 20% glucose aqueous solution;
sterilizing 900L LB culture medium at 121 deg.C under 0.101MPa for 20min, adding 100L sterilized 20% glucose aqueous solution, stirring, and mixing to obtain colony liquid culture medium.
12) Respectively inoculating strains of Lactobacillus plantarum, Pichia pastoris and Bacillus licheniformis in a colony liquid culture medium according to the inoculation amount of 1%, and culturing at room temperature overnight until OD of corresponding fermentation bacteria in each colony liquid culture medium600When the weight ratio of the fermentation broth to the fermentation broth reaches 1.5, the weight ratio of the fermentation broth to the fermentation broth is 1: 1: 1 to obtain a mixed bacterial preparation (marked as HJ 1). Wherein the inoculation amount is the weight content of the zymocyte in a colony liquid culture medium.
The fermentation inoculum prepared by the embodiment is used for preparing functional traditional Chinese medicine feed additives.
Example 2-9 preparation of fermentation inoculum
Embodiments 2 to 9 are respectively a method for preparing a fermentation inoculum, the steps of which are basically the same as those of embodiment 1, and the differences are only in the amount of raw materials and the process parameters, which are specifically shown in table 1:
TABLE 1 summary of the process parameters of examples 2 to 9
Example 10A method for preparing a functional Chinese medicinal feed additive
21) Taking 25kg of artemisia annua dregs sieved by a 40-mesh sieve, 5kg of naked flower purple bead powder, 5kg of folium artemisiae argyi dry leaves, 5kg of corn steep liquor dry powder, 10kg of soybean meal, 3kg of brown sugar and 2kg of stone powder to obtain a solid fermentation substrate (marked as M10).
22) Adding 45L mixed microbial inoculum HJ1 into solid fermentation substrate, wherein the total content of zymophyte is 2 hundred million CFU/g, mixing uniformly, and fermenting at room temperature for 12d to obtain functional Chinese medicinal feed additive (labeled as N10) with water content of 40%.
The functional traditional Chinese medicine feed additive prepared by the embodiment is suitable for being added into feed; the feed added with the functional traditional Chinese medicine feed additive is suitable for feeding livestock and poultry, in particular suitable for feeding livestock and poultry with enterovirus syndrome.
Example 11-15 preparation method of functional Chinese medicinal feed additive
Examples 11 to 15 are methods for preparing functional Chinese herbal feed additives, respectively, and the steps are substantially the same as those in example 10, except for differences in raw material amounts and process parameters, which are specifically shown in table 2:
TABLE 2 summary of the process parameters of examples 11 to 15
Wherein the fresh folium Artemisiae Argyi is obtained by picking, cleaning, and cutting.
The contents of the other portions of examples 11 to 15 are the same as those of example 10.
Example 16-21 preparation method of functional Chinese medicinal feed additive
Examples 16 to 21 are methods for preparing a functional Chinese herbal feed additive, which have substantially the same steps as in example 10, but differ only in the type and amount of raw materials and process parameters, as detailed in table 3:
TABLE 3 summary of the process parameters of examples 16 to 21
Wherein the fresh folium Artemisiae Argyi is obtained by picking, cleaning, and shearing.
The contents of the other portions of examples 16 to 21 are the same as those of example 10.
Example 22-27 preparation method of functional Chinese medicinal feed additive
Examples 22 to 27 are methods for preparing functional Chinese herbal feed additives, respectively, and the steps are substantially the same as in example 10, except for differences in the types, amounts, and process parameters of the raw materials, as detailed in table 4:
TABLE 4 summary of the process parameters of examples 22 to 27
Wherein the fresh folium Artemisiae Argyi is obtained by picking, cleaning, and shearing.
The contents of the other portions of examples 22 to 27 are the same as those of example 10.
Example 28-33 preparation method of functional Chinese medicinal feed additive
Examples 28 to 33 are methods for preparing a functional Chinese herbal feed additive, respectively, and the steps are substantially the same as in example 10, except for differences in the types, amounts, and process parameters of the raw materials, as detailed in table 5:
TABLE 5 summary of the process parameters of examples 28 to 33
Wherein the fresh folium Artemisiae Argyi is obtained by picking, cleaning, and shearing.
The contents of the other portions of examples 28 to 33 are the same as those of example 10.
Example 34-39 preparation method of functional Chinese medicinal feed additive
Examples 34 to 39 are methods for preparing a functional Chinese herbal feed additive, which have substantially the same steps as example 10, but differ only in the type and amount of raw materials and process parameters, as detailed in table 6:
TABLE 6 summary of the process parameters of examples 34 to 39
Wherein the fresh leaves of Artemisia annua and the fresh leaves of Artemisia argyi are respectively obtained by picking, cleaning and shearing.
The contents of the other portions of examples 34 to 39 are the same as those of example 10.
Example 40-45 preparation method of functional Chinese medicinal feed additive
Examples 40 to 45 are methods for preparing functional Chinese herbal feed additives, respectively, and the steps are substantially the same as those in example 10, except for differences in the types, amounts, and process parameters of the raw materials, as detailed in table 7:
TABLE 7 summary of the process parameters of examples 40 to 45
Wherein the fresh whole herb of Artemisia annua and fresh folium Artemisiae Argyi are obtained by picking, cleaning and shearing.
The contents of the other portions of examples 40 to 45 are the same as those of example 10.
Example 46-51 method for preparing functional Chinese medicinal feed additive
Examples 46 to 51 are methods for preparing a functional Chinese herbal feed additive, respectively, and the steps are substantially the same as those in example 10, except for differences in the types, amounts and process parameters of the raw materials, as detailed in table 8:
TABLE 8 summary of the process parameters of examples 46-51
Wherein the fresh stems and leaves of Artemisia annua are obtained by picking, cleaning and shearing.
The contents of the other portions of examples 46 to 51 are the same as those of example 10.
Example 52-57 preparation method of functional Chinese medicinal feed additive
Examples 52 to 57 are methods for preparing functional Chinese herbal feed additives, which have substantially the same steps as example 10, but differ only in the type and amount of raw materials and process parameters, as detailed in table 9:
TABLE 9 summary of the process parameters of examples 52 to 57
Wherein the fresh leaves, fresh whole plants, fresh stems and fresh leaves of Artemisia annua are respectively obtained by picking, cleaning and shearing.
The contents of the other portions of examples 52 to 57 are the same as those of example 10.
Example 58 content Change of protein, fat, cellulose and Secondary metabolite in functional Chinese medicinal feed additive
Respectively measuring crude protein content of objects to be measured (solid fermentation substrates M10-M16, M22, M28, M34, M40, M46, M52 and functional traditional Chinese medicine feed additives N10-N16, N22, N28, N34, N40, N46 and N52) by a biuret method, measuring fat content by referring to national standard, measuring cellulose content by referring to corresponding national standard, and measuring secondary metabolite content by a GC-MS method, wherein the secondary metabolite comprises dihydroartemisinic acid (A) and arteannuin (B). The specific measurement method is as follows, and the measurement results are shown in Table 11.
The content of crude protein is determined by a biuret method:
biuret reagent: collecting 1.5g copper sulfate (CuSO)4·5H2O) and 6.0g of sodium potassium tartrate are dissolved in 500mL of distilled water, 300mL of 10% sodium hydroxide aqueous solution is added while stirring, the volume is determined to be 1000mL, and the solution is stored in a reagent bottle coated with paraffin;
0.05 mol/L of the mixture was prepared-1Dissolving sodium hydroxide;
preparation of standard casein solution: accurately weighing 0.5g casein, dissolving in 0.05 mol.L-1The volume of the sodium hydroxide solution is determined to be 100mL, namely 5 mg/mL-1Preparing and measuring the standard solution of the complex protein according to the standard, and drawing a complex protein standard curve, wherein the ordinate is absorbance, and the abscissa is concentration, and the figure is 1.
And (3) sample determination:
drying the object to be measured at 80 ℃ to constant weight, and then placing the object in a dryer for cooling to obtain a dried sample for later use.
Weighing 0.2g of dried sample, putting the dried sample into a dry triangular flask, and adding 5mL of the dried sample with the concentration of 0.05 mol.L-1Wetting the sodium hydroxide solution, adding 20mL of biuret reagent, oscillating for 15min, standing at room temperature for reaction for 30min, filtering, carrying out color comparison on the obtained filtrate at the wavelength of 540nm, finding out the corresponding protein content on a standard curve, and adding 25mL of 10% sodium hydroxide solution as a blank control.
The crude fat content determination method is as follows:
the experimental process comprises the following steps: drying the extraction bottle in an oven at 105 +/-2 ℃ for 60min, cooling in a dryer for 30min, weighing, drying, cooling and weighing, and circulating the steps until the difference between the two weighing steps is less than 0.008g, wherein the weight is constant (the mass of the extraction bottle with the constant weight is obtained).
Drying the substance to be detected in a 105 ℃ oven for 120min, weighing 5g (drying to the mass of a sample to be detected with constant weight), pouring into an extraction bottle, adding 150ml of diethyl ether, heating in a 70 ℃ water bath until the diethyl ether flows back, controlling the reflux frequency of the diethyl ether to be about 10 times per hour, totally refluxing for 50 times, taking down the extraction bottle, evaporating residual diethyl ether on the water bath, wiping the outer wall of the extraction bottle, drying the extraction bottle in a 105 +/-2 ℃ oven for 120min to the constant weight, and weighing (the mass of the extraction bottle with constant weight and fat).
And (3) calculating: crude fat (%) = (the mass of an extraction bottle with constant weight + the mass of a sample to be tested after being dried to the constant weight-the mass of the extraction bottle with the fat with the constant weight)/the mass of a dried constant weight sample is 100 percent
Content of crude cellulose:
experimental reagent: the sulfuric acid solution has a concentration of 0.13 mol.L-1An aqueous solution of sulfuric acid; the sodium hydroxide solution had a concentration of 0.23 mol.L-1Aqueous sodium hydroxide solution.
(1) And (3) digestion of a sample: weighing 3g of sample to be detected, placing the sample to be detected in a 500mL reflux bottle, shaking uniformly, adding 150mL of sulfuric acid solution, and heating the sample on an electric heating furnace until the sample is boiled for 30 min;
(2) completely transferring the residue into a filter, and washing the residue with distilled water repeatedly for several times until the residual liquid is neutral;
(3) pouring the residue into a 500mL reflux bottle, adding 150mL sodium hydroxide solution, and continuously boiling for 30min to obtain residue X;
(4) taking a blank crucible, placing a small filter plate in the crucible, laying a layer of auxiliary materials, carefully pouring residue X, simultaneously performing suction filtration by using a vacuum pump, washing the residue X subjected to acid-base washing twice to be neutral by using water, washing for 3 times by using 30mL acetone solution, and draining filtrate after washing each time to obtain a residue sample;
(5) firing and drying: and (3) putting the crucible with the residue sample into a drying oven for drying at 130 ℃ for 2 hours, airing to room temperature, accurately weighing the mass of the crucible, putting the crucible into a muffle furnace for burning and ashing at 500 ℃ for 30min, and continuously twice.
(6) Blank experiment: the blank method was synchronized as described above, but the fermentation product was performed without the addition of bacteria.
And (3) determining the content of the secondary metabolite by a GC-MS method:
the chromatograph used was GCMS-QP20, the column HP-5 (30 mmx0.25mmx0.25mm) and the flow rate of carrier gas helium gas was 1.5mL min-1(ii) a The temperature of a sample inlet is 250 ℃, the split sample injection is carried out, and the split ratio is 1: 1, sampling amount is 1 mu L; the temperature of the column box is programmed to be 100 ℃, the temperature is kept for 1min, and then 6 ℃ min is carried out-1The temperature is raised to 220 ℃ at a high speedKept for 3min and then at 30 ℃ for min-1Heating to 280 ℃ at the speed of (1), and keeping for 5 min; the mass spectrum scanning range is 35-500 m/z. Wherein the GC-MS analysis determination chart measured by the functional Chinese medicinal feed additive N10 is shown in figure 2, wherein A is a general chart, 1 in A is dihydroartemisinic acid, 2 is artemisinic acid, 3 is artemisinine, 4 is artemisinin, B is a dihydroartemisinic acid analysis chart, C is an artemisinic acid analysis chart, D is an artemisinine analysis chart, and E is an artemisinin analysis chart.
TABLE 10 summary of the parameters of protein, fat, cellulose and secondary metabolites in solid fermentation substrate and functional Chinese medicinal feed additive
As can be seen from Table 10, the fermentation of Artemisia annua substances by using the fermentation inoculum reduces the cellulose content by 2.0-5.2%, increases the crude protein content by 5-16%, increases the fat content by more than 0.2%, and increases the four functional components in Artemisia annua, such as dihydroarteannuic acid, arteannuic acid, arteannuin and artemisinin after fermentation.
Example 59 application of functional Chinese medicinal feed additive in improving enterovirus syndrome of laying hens
Test one: application of functional traditional Chinese medicine feed additive in improvement of Lowman grey layer chicken enterovirus syndrome
a1) Test animals and design
The functional Chinese herbal medicine feed additives N10, N11, N16, N22, N28, N34, N40, N46 and N52 provided in examples 10, 11, 16, 22, 28, 34, 40, 46 and 52 are respectively added to the conventional commercial large layer chicken feed and the mixed feed is labeled as LN11 to LN110 in a one-to-one correspondence.
Selecting 12000 Roman grey laying hens with enterovirus syndrome at 240 days old, wherein the Roman grey laying hens have the symptoms of serious excessive feed and non-molding excrement, randomly dividing the Roman grey laying hens into 10 groups, 1200 laying hens in each group are respectively a control group and an A1~A9Group, wherein control group was fed with the currently marketed regular big layer feed, A1~A9Group-feeding laying hens respectivelyThe additive amount of the feed LN 11-LN 19 and the additive amount of N10, N11, N16, N22, N28, N34, N40, N46 and N52 are respectively 2.5 percent of the total amount of the layer chicken feed LN 11-LN 19. Feeding the feed all day long.
a2) Feeding management
The experiment is carried out on a large chicken farm in Cangzhou city or county in Hebei province. The experimental Roman gray laying hens are raised in three layers of three-dimensional cages, and are fed with dry powder, and freely fed and drunk with water. And (3) performing immunization and feeding management according to a conventional immunization program of the Roman grey laying hens, and sterilizing the henhouse every day to ensure the health of the henhouse. The feces and ingestion condition of the chicken flocks are observed every day during the test period. The test time is from 12 days 10 and 12 in 2019 to 24 days 10 and 24 in 2019.
a3) Measurement index and method
At the beginning of the test, the control group and A were taken separately1~A9Screening 1000g of feces of the group, wherein the screening feces data are shown in table 12, and after the test is finished, taking the control group and the group A again1~A91000g of feces were sieved, and the weight of the sieved feces is the total weight of the sieved feces, and the residue reduction rate = (total weight of sieved feces before test-total weight of sieved feces after test)/1000 g of feces are shown in table 12. Meanwhile, in the test period, the forming degree, the fineness, the eggshell gloss, the brightness and other data of the layer excrement are recorded by taking a group as a unit every day, see figure 3, the left side is the excrement of a control group at the end of the test, and the right side is A1The comparison of the excrement of the group shows that the excrement is shaped and becomes fine and smooth and the feed residue is obviously reduced when the feed added with the additive is fed. Referring to FIG. 4, eggs of the control group at the end of the test are shown on the left, and A is shown on the right1The comparison of the eggs of the group shows that the feed added with the additive of the invention obviously improves the glossiness of the eggshells, increases the brightness, and has no obvious change in the color and the eggshell quality.
TABLE 11 control and A1~A9Summary of the results of the group sifting manure weight
As can be seen from Table 11, after the feed added with the additive provided by the invention is fed, the excrement residue rate is obviously reduced, and the feed additive provided by the invention is proved to be capable of well improving the symptoms of serious overfeeding and unshaped excrement and relieving enterovirus syndrome of laying hens.
And (2) test II: application of functional traditional Chinese medicine feed additive in improvement of Lowman grey layer chicken enterovirus syndrome
b1) Test animals and design
The functional Chinese herbal medicine feed additives N10, N11, N16, N22, N28, N34, N40, N46 and N52 provided in examples 10, 11, 16, 22, 28, 34, 40, 46 and 52 are respectively added to the chicken feeds mixed with the existing commercial big layer chicken feeds and labeled LN21 to LN29 in a one-to-one correspondence manner.
Selecting 4800 Roman grey laying hens with enterovirus syndrome of 132 days old, wherein the Roman grey laying hens have symptoms of serious overfeeding and more red manure (suspected clostridium welchii infection), randomly dividing the Roman grey laying hens into 10 groups, and each group comprises 480 control groups and B1~B9Group, wherein control group was fed with the currently marketed feed for big layer hens, B1~B9The group is respectively fed with layer feed LN 21-LN 29, and the additive amounts of N10, N11, N16, N22, N28, N34, N40, N46 and N52 are respectively 5% of the total amount of the layer feed LN 21-LN 29. Feeding the feed all day long.
b2) Feeding management
The experiment was carried out on a large chicken farm in Cangzhou, Suanning county, Hebei province. The experimental Roman gray laying hens are raised in three layers of three-dimensional cages, and are fed with dry powder, and freely fed and drunk with water. And (3) performing immunization and feeding management according to a conventional immunization program of the Roman grey laying hens, and sterilizing the henhouse every day to ensure the health of the henhouse. The feces and ingestion condition of the chicken flocks are observed every day during the test period. The test time is from 12 days 4 and 12 months in 2019 to 24 days 4 and 24 months in 2019.
b3) Measurement index and method
At the beginning of the test, the control group and B were taken separately1~B9408g of feces of the group, screening the feces, and taking the control group and the B again after the test, wherein the screening feces data are shown in the table 131~B9408g of excrement, screening excrement, wherein the screening excrement data is shown in a table 13, wherein the weight of the screening excrement isThe residue reduction rate is = (total weight of screen dung before test-total weight of screen dung after test)/408 g for the total weight of screen with three layers. Meanwhile, in the test period, the data such as the red manure condition, the fineness and the like of the layer manure are recorded by taking groups as a unit every day, and the test data show that when the additive is fed to the feed added with the additive, the red manure condition is obviously improved and becomes fine and smooth, and the feed residue is obviously reduced.
TABLE 12 control and B1~B9Summary of the results of the group sifting manure weight
As can be seen from Table 12, after the feed added with the additive provided by the invention is fed, the excrement residue rate is obviously reduced, and the feed additive provided by the invention is proved to be capable of well improving the symptoms of serious overfeeding and more red excrement and relieving enterovirus syndrome of laying hens.
And (3) test III: application of functional traditional Chinese medicine feed additive in improvement of weaned piglet enterovirus syndrome
c1) Test animals and design
The functional Chinese medicinal feed additives N10, N11, N16, N22, N28, N34, N40, N46 and N52 provided in examples 10, 11, 16, 22, 28, 34, 40, 46 and 52 are respectively added to the pig feed mixed with the existing commercial pig feed to mark LN31 to LN39 in a one-to-one correspondence manner.
Selecting 120 weaned piglets with age of 28 days, wherein the weaned piglets have symptoms (suspected enterotoxigenic lesions) such as constipation, round and round belly, unbearable abdominal pain of pigs, restlessness, shaking head and tail, and limb after lying on stomach for defecation and urination, but do not discharge fecaluria, shortness of breath, and white foam in mouth, and randomly dividing into 10 groups, wherein each group comprises 12 weaned piglets comprises a control group and C1~C9Group, in which the control group was fed with the currently marketed pig feed, C1~C9The feed is respectively fed to pig feed LN 31-LN 39, and the addition amounts of additives N10, N11, N16, N22, N28, N34, N40, N46 and N52 are respectively 5% of the total amount of the pig feed LN 31-LN 39. Feeding three days per day, and culturing for 40 days.
c2) Feeding management
The experiment is carried out in a large farm in Cangzhou city of Hebei province. Pigs were fed and drunk freely. The pigsty adopts natural ventilation and mechanical ventilation and a cement floor. According to the normal feeding management of a pig farm, the pig feeding, the excrement and the mental conditions are observed every day, and diseases are found and recorded and processed in time.
c3) Measurement index and method
Weaned piglets were weighed on an empty stomach at the beginning and end of the trial, respectively. The average daily gain of the pigs was calculated over 50 days of feeding and the diarrhea status of the weaned piglets was recorded. The specific test results are as follows:
table 13 summary of the application effect of the functional feed additive in weaned pig breeding
As can be seen from Table 13, the diarrhea and death of the weaned piglets are reduced and the daily gain is increased after the feed added with the additive is fed. The feed additive disclosed by the invention is proved to be capable of well relieving enterovirus syndrome of weaned piglets.
It should be noted that, the embodiments 1 to 59 are only preferred embodiments of the present invention, and are not intended to limit the present invention in other forms, and especially, the fermentation bacteria agent used in the fermentation process is not limited to the specific bacteria agent used in the embodiments 10 to 57, and any person skilled in the art may use the above technical content as a teaching to modify or modify the above embodiments into equivalent embodiments with equivalent changes, but all the simple modifications, equivalent changes and modifications made to the above embodiments without departing from the technical spirit of the present invention, and still fall within the scope of the present invention as claimed.
Claims (8)
1. A functional Chinese medicinal feed additive is characterized in that raw materials for preparing effective components of the feed additive comprise artemisia annua substances, auxiliary materials and zymophyte; wherein the dosage of the raw materials adopts any one scheme or the combination of at least two schemes from a) to g),
a) the artemisia annua substances are 20-30 parts of artemisia annua dregs;
the auxiliary materials comprise: 3-8 parts of naked flower purple bead powder, 3-8 parts of dried folium artemisiae argyi/6-16 parts of fresh folium artemisiae argyi, 3-8 parts of dried corn steep liquor/6-16 parts of corn steep liquor, 8-12 parts of soybean meal, 2-4 parts of brown sugar and 1-3 parts of mountain flour;
the total amount of the zymophyte agent is more than or equal to 10 hundred million CFU/g;
the water content of the raw material is 30-50%;
b) the artemisia annua substances are 10-20 parts of artemisia annua dry leaves in parts by weight;
the auxiliary materials comprise: 3-8 parts of naked flower purple bead powder, 3-8 parts of dried folium artemisiae argyi/6-16 parts of fresh folium artemisiae argyi, 3-8 parts of dried corn steep liquor/6-16 parts of corn steep liquor, 8-12 parts of soybean meal, 2-4 parts of brown sugar and 1-3 parts of mountain flour;
the total amount of the zymophyte agent is more than or equal to 10 hundred million CFU/g;
the water content of the raw material is 30-50%;
c) the artemisia annua substances are 30-50 parts of artemisia annua dry whole plants in parts by weight;
the auxiliary materials comprise: 3-8 parts of naked flower purple bead powder, 3-8 parts of dried folium artemisiae argyi/6-16 parts of fresh folium artemisiae argyi, 3-8 parts of dried corn steep liquor/6-16 parts of corn steep liquor, 8-12 parts of soybean meal, 2-4 parts of brown sugar and 1-3 parts of mountain flour;
the total amount of the zymophyte agent is more than or equal to 10 hundred million CFU/g;
the water content of the raw material is 30-50%;
d) the artemisia annua substances are 50-70 parts of artemisia annua dry stems by weight;
the auxiliary materials comprise: 3-8 parts of naked flower purple bead powder, 3-8 parts of dried folium artemisiae argyi/6-16 parts of fresh folium artemisiae argyi, 3-8 parts of dried corn steep liquor/6-16 parts of corn steep liquor, 8-12 parts of soybean meal, 2-4 parts of brown sugar and 1-3 parts of mountain flour;
the total amount of the zymophyte agent is more than or equal to 10 hundred million CFU/g;
the water content of the raw material is 30-50%;
e) the artemisia annua substances are 40-60 parts of artemisia annua fresh leaves in parts by weight;
the auxiliary materials comprise: 3-8 parts of naked flower purple bead powder, 3-8 parts of dried folium artemisiae argyi/6-16 parts of fresh folium artemisiae argyi, 3-8 parts of dried corn steep liquor/6-16 parts of corn steep liquor, 8-12 parts of soybean meal, 2-4 parts of brown sugar and 1-3 parts of mountain flour;
the total amount of the zymophyte agent is more than or equal to 10 hundred million CFU/g;
the water content of the raw material is 30-50%;
f) the artemisia annua substances are 50-70 parts of fresh artemisia annua whole plants in parts by weight;
the auxiliary materials comprise: 3-8 parts of naked flower purple bead powder, 3-8 parts of dried folium artemisiae argyi/6-16 parts of fresh folium artemisiae argyi, 3-8 parts of dried corn steep liquor/6-16 parts of corn steep liquor, 8-12 parts of soybean meal, 2-4 parts of brown sugar and 1-3 parts of mountain flour;
the total amount of the zymophyte agent is more than or equal to 10 hundred million CFU/g;
the water content of the raw material is 30-50%;
g) 60-80 parts by weight of fresh stems of artemisia annua;
the auxiliary materials comprise: 3-8 parts of naked flower purple bead powder, 3-8 parts of dried folium artemisiae argyi/6-16 parts of fresh folium artemisiae argyi, 3-8 parts of dried corn steep liquor/6-16 parts of corn steep liquor, 8-12 parts of soybean meal, 2-4 parts of brown sugar and 1-3 parts of mountain flour;
the total amount of the zymophyte agent is more than or equal to 10 hundred million CFU/g;
the water content of the raw material was 40%.
2. The functional Chinese herbal medicine feed additive according to claim 1, wherein,
the fermentation microbial inoculum comprises at least one of lactobacillus plantarum, pichia pastoris, bacillus licheniformis, aspergillus niger, lactobacillus casei, trichoderma reesei and bacillus subtilis.
3. The preparation method of the functional Chinese herbal medicine feed additive according to claim 1 or 2, characterized in that the artemisia annua is taken and added with auxiliary materials and fermentation bacteria agent to be mixed and fermented, and the functional Chinese herbal medicine feed additive is obtained.
4. The method for preparing functional Chinese medicinal feed additive according to claim 3,
sieving the artemisia annua dregs, the artemisia annua dry leaves, the artemisia annua dry whole grass, the artemisia annua dry stems and stems, the callicarpa nudiflora powder, the artemisia argyi dry leaves, the corn steep liquor dry powder, the bean pulp, the brown sugar and the stone powder to obtain a granularity of more than or equal to 40 meshes;
the fresh leaves, fresh whole plants, fresh stems and stems of Artemisia annua and fresh leaves of Artemisia argyi are respectively cut leaves, whole plants, stems and stems of Artemisia annua and Artemisia argyi.
5. The preparation method of the functional traditional Chinese medicine feed additive according to claim 3, wherein the fermentation temperature is room temperature and the fermentation time is 12-30 days.
6. The method for preparing the functional Chinese medicinal feed additive according to any one of claims 3 to 5, wherein when the fermentation inoculum is a mixed inoculum, the fermentation inoculum is inoculated to the culture medium and cultured to OD600After the value is 1.2-1.8, mixing fermentation liquor corresponding to the obtained fermentation strain to obtain the fermentation strain;
when the zymophyte agent is a single agent, a zymophyte is taken and inoculated in a culture medium to be cultured to OD600And obtaining the product after the value is 1.2-1.8.
7. The use of the functional Chinese herbal medicine feed additive of claim 1 or 2, wherein the functional Chinese herbal medicine feed additive is used for being added into feed for feeding livestock and poultry.
8. The use of the functional Chinese medicinal feed additive according to claim 7, wherein the feed containing the functional Chinese medicinal feed additive is used for relieving enterovirus syndrome of livestock and poultry.
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Application publication date: 20201023 |