CN112704150A - Fermented traditional Chinese medicine immunopotentiator and preparation method and application thereof - Google Patents
Fermented traditional Chinese medicine immunopotentiator and preparation method and application thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/10—Feeding-stuffs specially adapted for particular animals for ruminants
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- 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
- A23K50/60—Feeding-stuffs specially adapted for particular animals for weanlings
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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)
- A61K36/31—Brassicaceae or Cruciferae (Mustard family), e.g. broccoli, cabbage or kohlrabi
- A61K36/315—Isatis, e.g. Dyer's woad
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- 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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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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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
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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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
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
Abstract
The invention provides a fermented traditional Chinese medicine immunopotentiator, and belongs to the technical field of feed additives. The fermented traditional Chinese medicine immunopotentiator provided by the invention is obtained by carrying out water extraction on radix isatidis and radix astragali and then fermenting. The isatis root and the astragalus are subjected to water extraction and fermentation, and the fermented traditional Chinese medicine not only improves the content of active ingredients of the medicine, but also contains probiotics beneficial to the health of animals. The results of the examples show that: the traditional Chinese medicine immunopotentiator provided by the invention obviously improves the polysaccharide content in the medicine, improves the abundance and diversity of intestinal flora of lambs after the fermented traditional Chinese medicine immunopotentiator is fed to weaned lambs, obviously improves the immunity of weaned lambs, reduces the natural infection rate of coccidian, promotes the growth of sheep, improves the daily gain and reduces the material-weight ratio.
Description
Technical Field
The invention belongs to the technical field of feed additives, and particularly relates to a fermented traditional Chinese medicine immunopotentiator and a preparation method and application thereof.
Background
The weaning of the lambs has various stress reactions, the weaning lambs in the stage are often low in immunity and easy to be infected with various pathogens, and the weaning lambs are generally prevented by clinically using antibiotics. However, when killing pathogenic bacteria, antibiotics can cause drug residues, cause drug resistance of bacteria, imbalance of animal intestinal flora and other problems, and seriously affect the healthy development of animal husbandry and harm public health safety. More and more countries forbid antibiotic products as animal growth promoters, and China also successively promulgates a series of laws and regulations to forbid the use of growth-promoting antibiotics in feed. Therefore, the development of new antibiotic substitutes that can both enhance resistance and promote growth in animals is urgent.
Disclosure of Invention
In order to solve the problems, the invention provides a fermented traditional Chinese medicine immunopotentiator and a preparation method and application thereof. The fermented traditional Chinese medicine immunopotentiator provided by the invention can obviously improve the immunity of organisms and replace antibiotics.
In order to achieve the above purpose, the invention provides the following technical scheme:
the invention provides a fermented traditional Chinese medicine immunopotentiator, which is obtained by carrying out water extraction on isatis root and astragalus and then fermenting.
Preferably, the fermenting bacteria comprise bacillus subtilis.
The invention provides a preparation method of a traditional Chinese medicine immunopotentiator, which comprises the following steps: carrying out water extraction on the isatis root and the astragalus to obtain an aqueous extract, and mixing the aqueous extract with an LB basal culture medium to obtain a fermentation culture medium; inoculating zymophyte into the fermentation culture medium for fermentation to obtain the fermented traditional Chinese medicine immunopotentiator.
Preferably, the aqueous extraction comprises the steps of: soaking radix astragali and radix Isatidis in water, decocting, filtering, and concentrating the filtrate to obtain water extract.
Preferably, the mass ratio of the astragalus to the isatis root is (1-5): 1; the volume ratio of the total mass of the astragalus and the isatis root to the water added each time is 1 g: (3-20) mL; the soaking time is 20-60 min; the decocting temperature is 90-100 ℃, and the decocting time is 30-60 min; the number of times of decoction is 1-3.
Preferably, the volume ratio of the aqueous extract to the LB basic medium is 1: (1-5).
Preferably, the LB basic culture medium comprises the following components in mass concentration: 1-10 g/L of peptone, 10-30 g/L of yeast extract powder, 10-50 g/L of glucose and MnSO4·7H2O1-3 g/L; the pH value of the LB basic culture medium is 7.0-7.2.
Preferably, the inoculation amount of the zymophyte is 1-10%.
Preferably, the fermentation temperature is 30-40 ℃, and the fermentation time is 24-48 h.
The invention provides the fermented traditional Chinese medicine immunopotentiator in the technical scheme or the application of the traditional Chinese medicine immunopotentiator obtained by the preparation method in the technical scheme in livestock breeding.
Has the advantages that:
the invention provides a fermented traditional Chinese medicine immunopotentiator, which is obtained by carrying out water extraction on isatis root and astragalus and then fermenting. The isatis root and the astragalus are subjected to water extraction and fermentation, the fermented traditional Chinese medicine not only improves the efficacy, remarkably improves the immunity of the organism and the richness and diversity of the flora in duodenum, and relieves various stress reactions of the sheep in the weaning period, but also has the advantages of low residue and low toxic and side effects, and can be used for replacing antibiotics. The results of the examples show that: the fermented traditional Chinese medicine immunopotentiator provided by the invention obviously improves the polysaccharide content, and after the fermented traditional Chinese medicine immunopotentiator is fed to the weaned sheep, the immunity of the sheep is obviously improved, the growth of the sheep is promoted, and the stress reaction of the weaned sheep is obviously relieved.
Drawings
FIG. 1 is a glucose standard curve of example 5.
Detailed Description
The invention provides a fermented traditional Chinese medicine immunopotentiator, which is obtained by carrying out water extraction and fermentation on isatis root and astragalus. In the invention, the sources of the isatis root and the astragalus root are not specially required, and the products which are conventional and commercially available in the field can be adopted. In the invention, the preferable radix isatidis and the preferable radix astragali are radix isatidis decoction pieces and radix astragali decoction pieces respectively, and the radix isatidis decoction pieces and the radix astragali decoction pieces are prepared from common commercially available products. In the invention, the fermented zymophyte preferably comprises bacillus subtilis, the bacillus subtilis is preferably bacillus subtilis with the number of Y5 which is separated, identified and stored in a microbiological laboratory of the institute of medicine and engineering of Henan agricultural university, the bacillus is an existing strain, and references are as follows: zhanghongying, et al, determination of antibacterial activity and cellulase activity of Bacillus, J. Microecology, Vol. 21, No. 9, 2009. The bacillus subtilis has good enzymes for synthesizing alpha-amylase, protease, lipase, cellulase and the like, can improve the expression level of rumen epithelial cell beta-defensin, and improves the disease resistance of organisms. The isatis root and the astragalus are subjected to water extraction and fermentation, and the fermented traditional Chinese medicine not only improves the efficacy, but also obviously improves the immunity of the organism and relieves various stress reactions of the sheep in the weaning period; the radix isatidis and the astragalus membranaceus have the advantages of high curative effect, low residue and low toxic and side effects after fermentation, and can be used for replacing antibiotics.
The invention provides a preparation method of a fermented traditional Chinese medicine immunopotentiator, which comprises the following steps: carrying out water extraction on the isatis root and the astragalus to obtain an aqueous extract, and mixing the aqueous extract with an LB basal culture medium to obtain a fermentation culture medium; inoculating zymophyte into the fermentation culture medium for fermentation to obtain the fermented traditional Chinese medicine immunopotentiator.
The invention carries out water extraction on the isatis root and the astragalus to obtain a water extract. In the present invention, the aqueous extraction preferably comprises the steps of: soaking radix astragali and radix Isatidis in water, decocting, filtering, and concentrating the filtrate to obtain water extract. In the invention, the mass ratio of the astragalus to the isatis root is preferably (1-5): 1, more preferably 1: 2; in the invention, the volume ratio of the total mass of the astragalus and the isatis root to each water addition is preferably 1 g: (3-20) mL, more preferably 1 g: 5 mL. In the invention, the soaking time is preferably 20-60 min, and more preferably 30min, so that effective components in the traditional Chinese medicine can be fully dissolved out by soaking, and the utilization rate of the traditional Chinese medicine is improved. In the invention, the decoction temperature is preferably 90-100 ℃, and more preferably 100 ℃; in the invention, the decoction time is preferably 30-60 min, and more preferably 40 min; in the present invention, the number of times of decoction is preferably 1 to 3 times, and more preferably 3 times. The invention selects specific extraction conditions to fully dissolve out the effective substances in the isatis root and the astragalus root and improve the utilization rate of medicinal materials. After decocting, the filtrate obtained by filtering after decocting is preferably concentrated to obtain an aqueous extract in the invention. In the present invention, the concentration method is preferably concentration by a rotary evaporator, the concentration of the crude drug in the concentrated drug solution is preferably 1g/mL, and in the present invention, the concentrated drug solution is preferably autoclaved to obtain the aqueous extract.
After obtaining the aqueous extract, the invention mixes the aqueous extract with LB basic culture medium to obtain the fermentation culture medium. In the present invention, the volume ratio of the aqueous extract to the LB basal medium is preferably 1: (1-5), and more preferably 1: 3. In the present invention, the LB basal medium preferably comprises the following components in mass concentrations: 1-10 g/L of peptone, 10-30 g/L of yeast extract powder, 10-50 g/L of glucose and MnSO4·7H2O1-3 g/L; (ii) a Further preferably comprises: 5g/L of peptone, 30g/L of yeast extract powder, 20g/L of glucose and MnSO4·7H2O2 g/L. In the invention, the pH of the LB basic culture medium is preferably 7.0-7.2, and more preferably 7.2.
After the fermentation culture medium is obtained, the invention inoculates zymophyte into the fermentation culture medium for fermentation to obtain the fermented traditional Chinese medicine immunopotentiator. In the present invention, the amount of the fermentation tubes inoculated is preferably 1% to 10%, more preferably 5%, and the number of viable bacteria of the fermentation tubes in the fermentation medium after inoculation is preferably 3 × 107～3*1010CFU/mL, more preferably 3X 109CFU/mL. In the invention, the fermentation temperature is preferably 30-40 ℃, and more preferably 35 ℃; the fermentation time is preferably 24-48 h, and more preferably 36 h.
The preparation method of the fermented traditional Chinese medicine immunopotentiator provided by the invention is simple in process, the fermentation process is easy to control, and the prepared fermented traditional Chinese medicine immunopotentiator has the advantages of high curative effect, low residue and low toxic and side effects.
The invention provides an application of the fermented traditional Chinese medicine immunopotentiator in the technical scheme or the fermented traditional Chinese medicine immunopotentiator obtained by the preparation method in the technical scheme in livestock breeding. In the invention, the fermented traditional Chinese medicine immunopotentiator is preferably added into a basic feed in a mode of mixing materials for feeding. In the invention, the dosage of the fermented traditional Chinese medicine immunopotentiator is preferably 2-8 g/d.
In order to further illustrate the present invention, the following examples are provided to describe the fermented traditional Chinese medicine immunopotentiator and the preparation method and application thereof in detail, but they should not be construed as limiting the scope of the present invention.
Example 1
A fermented traditional Chinese medicine immunopotentiator is prepared by the following steps:
1) preparation of water extract of radix Isatidis and radix astragali
Accurately weighing 1000g of astragalus decoction pieces and 1000g of isatis root decoction pieces by using a balance; mixing radix astragali and radix Isatidis, soaking in water at a volume ratio of 1g/5mL to total mass of radix astragali and radix Isatidis, soaking for 30min, decocting at 100 deg.C for 40min, filtering with gauze, adding distilled water into residue, decocting for 40min, and repeatedly decocting for 3 times; decocting for 3 times, filtering with gauze to obtain medicinal liquid, placing into rotary evaporator, concentrating the medicinal liquid to obtain medicinal liquid containing crude drug 1g/mL, and autoclaving to obtain radix Isatidis and radix astragali water extractive solution.
2) Fermentation of
And taking 250mL of the water extract of the radix isatidis and the radix astragali, and then adding the water extract of the radix isatidis and the radix astragali into 750mL of LB basal medium to obtain a fermentation medium. Wherein the LB basic culture medium consists of the following components: peptone: 5g/L, yeast extract powder: 30g/L, glucose: 20g/L, MnSO4 & 7H 2O: 2g/L, pH: 7.2. after obtaining the fermentation medium, inoculating bacillus subtilis (separated, identified and stored in a microbiological laboratory of institute of pharmaceutical engineering, university of Henan agriculture) into the fermentation medium, wherein the inoculation amount of the fermentation bacteria is 5 percent (volume ratio), and after inoculation, the viable count of the fermentation bacteria in the fermentation medium is 3 x 109CFU/mL, the fermentation temperature is 35 ℃, and the fermentation time is 36h, so as to obtain the traditional Chinese medicine immunopotentiator.
Example 2
A fermented traditional Chinese medicine immunopotentiator is prepared by the same method as the example 1, and the difference is that: the fermentation temperature is 30 ℃, and the fermentation time is 48 h.
Example 3
A fermented traditional Chinese medicine immunopotentiator is prepared by the same method as the example 1, and the difference is that: the fermentation temperature is 37 ℃, and the fermentation time is 24 h.
Example 4
A fermented traditional Chinese medicine immunopotentiator is prepared by the same method as the example 1, and the difference is that: the fermentation temperature is 30 ℃, and the fermentation time is 48 h.
Comparative example 1
A fermented traditional Chinese medicine immunopotentiator is prepared by the same method as the example 1, and the difference is that: no LB medium was added during fermentation.
Example 5
The change of polysaccharide content in fermentation liquid of radix Isatidis and radix astragali before and after fermentation
The change of the polysaccharide content in the fermentation liquid of the radix isatidis and the radix astragali before and after fermentation in examples 1-4 and comparative example 1 is examined, and the influence of the fermentation on the polysaccharide content is examined.
The detection method comprises the following steps: precisely weighing 10mg of glucose standard product dried to constant weight at 105 ℃, placing the glucose standard product in a 100mL volumetric flask, adding water to dissolve the glucose standard product, diluting the glucose standard product to a scale, and shaking up to obtain 0.1mg of standard solution containing glucose standard product per 1 mL. 0.2g of anthrone (with the purity of 99.99%) is weighed, added with 100mL of concentrated sulfuric acid, and placed in a brown reagent bottle and a refrigerator for later use. 0mL, 0.1mL, 0.4mL, 0.6mL, 0.8mL, 1.0mL, 1.2mL, 1.4mL, 1.6mL of the standard solution of 0.1mg/mL was precisely aspirated into 10mL stoppered tubes, and then 2mL of the standard solution was added with distilled water, and the samples were numbered. Then adding 6mL of prepared anthrone reagent, shaking up, heating in boiling water bath for 15min, taking out, rapidly cooling in ice water to room temperature, measuring absorbance at wavelength of 620nm with anthrone reagent as blank, repeating for multiple times to obtain regression equation, and glucose standard curve is shown in FIG. 1.
Centrifuging 3mL of water extract before and after fermentation at 4000r/min respectively, centrifuging for 10min, adding 4mL of absolute ethyl alcohol into 1mL of supernate, standing for 2-3 h, and centrifuging again; washing the precipitate with 95% ethanol for 2 times, each time 5mL, heating distilled water 2mL to dissolve the precipitate, fixing the volume to 100mL, shaking, placing 0.5mL solution in a 10mL volumetric flask, adding water to fix the volume to the scale, and mixing to obtain the test solution. And (3) putting the sample to be measured into a photometer to measure the absorbance, and repeatedly taking the absorbance into a regression equation to obtain the polysaccharide content.
TABLE 1 polysaccharide content in fermentation broth before and after fermentation
As can be seen from the results in table 1, the contents of polysaccharides in the fermented solutions after fermentation in examples 1 to 4 were significantly higher than those before fermentation, and it was found that the fermentation had the effect of increasing the polysaccharide content in the fermented solution, thereby improving the immunity of animals. The change of polysaccharide content before and after fermentation in comparative example 1 was not large, and it is probably because no LB medium was added to the fermentation broth, so that Bacillus subtilis could not proliferate and the fermentation effect could not be achieved.
Example 6
Effect on sheep growth Performance
Test time: 11-12 months in 2019;
test site: a three-wood green source livestock breeding base in a sealing city;
sheep species: 40 days old Hu sheep;
selecting 18 sheep, randomly dividing into 3 groups, namely a control group, a fermented group and an unfermented group, wherein the control group is fed with basic feed only and is used for collecting water normally; the fermentation group feeds the traditional Chinese medicine reinforcing agent and the basic feed fermented in the example 1, the feeding amount of the traditional Chinese medicine reinforcing agent is 6 g/feed/d, and the traditional Chinese medicine reinforcing agent is fed by mixing with the feed every day; the unfermented group is fed with the traditional Chinese medicine reinforcing agent and the basal feed before fermentation in example 1, the feeding amount of the traditional Chinese medicine reinforcing agent is 6 g/per day, and the traditional Chinese medicine reinforcing agent is fed with the feed every day for 28 days. The feed intake and weight of each group of sheep are recorded every day, the influence of the traditional Chinese medicine reinforcing agent on the growth performance of the sheep is examined, and the result is shown in table 2.
TABLE 2 Effect of the present invention fermented traditional Chinese medicine enhancers on sheep growth Performance
| Item | Average daily food intake (g/d) | Average daily gain (g/d) | Material to weight ratio |
| Control group | 851.79 | 118.57±28.73 | 5.96 |
| Fermentation group | 914.29 | 208.57±42.68** | 4.38 |
| Non-fermented group | 821.43 | 145.71±35.21* | 5.64 |
Note: indicates a very significant difference from the blank control group (P <0.01) and indicates a significant difference from the blank control group (P < 0.05).
As can be seen from Table 2, the average daily gain of the fermented group is significantly higher than that of the control group and also significantly higher than that of the drug unfermented group, and the material weight ratio of the fermented group to the fermented group is the lowest.
Example 7
Influence on sheep immune function
Test time: 11-12 months in 2019;
test site: a three-wood green source livestock breeding base in a sealing city;
sheep species: 40 days old Hu sheep;
selecting 18 sheep, randomly dividing into 3 groups, namely a control group, a fermented group and an unfermented group, wherein the control group is fed with basic feed only and is used for collecting water normally; the fermentation group feeds the traditional Chinese medicine immunopotentiator and the basic feed fermented in the example 5, the feeding amount of the traditional Chinese medicine immunopotentiator is 6 g/d, and the traditional Chinese medicine immunopotentiator is fed by mixing with the feed every day; the unfermented group is fed with the traditional Chinese medicine immunopotentiator and the basal feed before fermentation in example 1, the feeding amount of the traditional Chinese medicine immunopotentiator is 6 g/d, and the traditional Chinese medicine immunopotentiator is fed with the feed every day. The sheep were fed for 28 days according to the above test protocol, and 5 sheep were randomly selected from each group per week for blood sampling, and the immune-related index in the sheep serum was determined (ELISA kit, Shanghai FANKEL Industrial Co. Ltd., brand: Fankew). The results are shown in Table 3.
TABLE 3 Effect of the Chinese medicinal immunopotentiator of the present invention on sheep's immune function
Note: indicates significant difference compared to the blank control group and the unfermented group (P < 0.05); -not detected.
The results in table 3 show that the immune globulin and interleukin in the 7d, 14d, 21d and 28d sheep and the fermentation group are obviously increased compared with the blank control and the unfermented group, which indicates that the fermented traditional Chinese medicine prepared by the invention can enhance the immunity of sheep organisms.
Example 8
Effect on sheep intestinal flora
Test time: 11-12 months in 2019;
test site: a three-wood green source livestock breeding base in a sealing city;
sheep species: 40 days old Hu sheep;
selecting 18 sheep, randomly dividing into 3 groups, namely a control group, a fermented group and an unfermented group, wherein the control group is fed with basic feed only and is used for collecting water normally; the fermentation group feeds the traditional Chinese medicine immunopotentiator and the basic feed fermented in the example 1, the feeding amount of the traditional Chinese medicine immunopotentiator is 6 g/d, and the traditional Chinese medicine immunopotentiator is fed by mixing with the feed every day; the unfermented group is fed with the traditional Chinese medicine immunopotentiator and the basal feed before fermentation in example 1, the feeding amount of the traditional Chinese medicine immunopotentiator is 6 g/d, and the traditional Chinese medicine immunopotentiator is fed with the feed every day. Feeding for 28d according to the test scheme, killing and collecting duodenum, jejunum and ileum sections after the test is finished, determining DNA sequences of sheep intestinal flora microorganisms by adopting Illumina high-throughput sequencing technology for collected intestinal section contents (generation determination of Pesenno Biotech Co., Ltd.), and comprehensively analyzing diversity and abundance of intestinal microflora. The results are shown in Table 4.
TABLE 4 influence of fermented traditional Chinese medicine feed on alpha diversity of sheep duodenum content
| Index of refraction | Fermentation group | Non-fermented group | Control group |
| Chao1 | 2268.92a | 1756.75ab | 985.01b |
| Shannon | 6.94a | 5.27a | 3.73b |
Note: letter difference indicates significant difference (P <0.05)
The total of 36 phyla, 100 class, 190 order, 315 family and 597 genera of bacteria were determined by OTU clustering and species annotation in all samples in this experiment.
The series of statistical analysis indices allow assessment of species abundance and diversity of bacterial communities in a sample, where a-diversity reflects the diversity of species within a single sample, including species observation index (observedspececiesendex), super 1 index Chao 1 index, eis index (Ace index), Shannon index (Shannon index) and simpson index (Simpsonindex). Wherein the species observation index and the Chao 1 index reflect the abundance of the community (specificesichness) in the sample, the Shannon index and the Simpson index reflect the diversity of the community. The experiment evaluates the abundance and diversity of microbial communities primarily by the Chao 1 index and the shannon index. Post-hoc tests (table 4) with dunn' test as pairwise comparison show that the Chao 1 index and the shannon index of the fermentation group are significantly higher than those of the blank control group (P <0.05), which indicates that the bacillus subtilis fermentation astragalus extract has significant influence on the abundance and diversity of the sheep duodenum flora. The abundance and diversity of the flora in the jejunum and ileum were not significantly different (data not shown).
The results of the above examples show that the fermented traditional Chinese medicine immunopotentiator provided by the invention significantly increases the polysaccharide content, and after the fermented traditional Chinese medicine immunopotentiator provided by the invention is fed to weaned sheep, the sheep immunity is significantly increased, the abundance and diversity of the duodenum flora are significantly increased, and the growth of the weaned sheep is promoted.
Although the present invention has been described in detail with reference to the above embodiments, it is only a part of the embodiments of the present invention, not all of the embodiments, and other embodiments can be obtained without inventive step according to the embodiments, and the embodiments are within the scope of the present invention.
Claims (10)
1. A fermented traditional Chinese medicine immunopotentiator is characterized in that the fermented traditional Chinese medicine immunopotentiator is obtained by carrying out water extraction on radix isatidis and radix astragali and then fermenting.
2. The fermented traditional Chinese medicine immunopotentiator of claim 1, wherein the fermented zymophyte comprises bacillus subtilis.
3. The method for preparing the fermented traditional Chinese medicine immunopotentiator according to claim 1 or 2, comprising the steps of: carrying out water extraction on the isatis root and the astragalus to obtain an aqueous extract, and mixing the aqueous extract with an LB basal culture medium to obtain a fermentation culture medium; inoculating zymophyte into the fermentation culture medium for fermentation to obtain the fermented traditional Chinese medicine immunopotentiator.
4. The method for preparing according to claim 3, wherein said aqueous extraction comprises the steps of: soaking radix astragali and radix Isatidis in water, decocting, filtering, and concentrating the filtrate to obtain water extract.
5. The preparation method according to claim 4, wherein the mass ratio of the astragalus to the isatis root is (1-5): 1; the volume ratio of the total mass of the astragalus and the isatis root to the water added each time is 1 g: (3-20) mL; the soaking time is 20-60 min; the decocting temperature is 90-100 ℃, and the decocting time is 30-60 min; the number of times of decoction is 1-3.
6. The method of claim 3, wherein the volume ratio of the aqueous extract to LB basal medium is 1: (1-5).
7. The method according to claim 3, wherein the LB basal medium comprises the following components in mass concentration: 1-10 g/L of peptone, 10-30 g/L of yeast extract powder, 10-50 g/L of glucose and MnSO4·7H2O1-3 g/L; the pH value of the LB basic culture medium is 7.0-7.2.
8. The method according to claim 3, wherein the amount of the inoculated fermentation tubes is 1% to 10%.
9. The method according to claim 3, wherein the temperature of the fermentation is 30-40 ℃ and the time of the fermentation is 24-48 h.
10. The fermented traditional Chinese medicine immunopotentiator according to claim 1 or 2 or the fermented traditional Chinese medicine immunopotentiator obtained by the preparation method according to any one of claims 3 to 9, for use in livestock breeding.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1279727A4 (en) * | 2000-04-26 | 2004-09-01 | Bingxin Wu | Transformation and modification of natural drug by intestinal probiotic fermentative culture |
| CN101099536A (en) * | 2006-07-03 | 2008-01-09 | 河南农业大学 | Application of embedded lactobacillus in animal feed and bacteriostatic promoting production |
| CN106591182A (en) * | 2016-12-08 | 2017-04-26 | 王丽娟 | Microecological preparation with bacillus subtilis fermenting astragalus membranaceus and application thereof |
| CN106666247A (en) * | 2017-03-23 | 2017-05-17 | 威海金牌生物科技股份有限公司 | Foraging feed of special compound feed for breeding stichopus japonicus and preparation method of foraging feed |
| CN106880777A (en) * | 2017-02-27 | 2017-06-23 | 武文海 | A kind of Helth-care product with functions of prolonging life and nourishing face and preparation method thereof |
| CN107319546A (en) * | 2017-08-07 | 2017-11-07 | 黑龙江省兽医科学研究所 | A kind of fermented tcm powder and preparation method thereof |
| EP2914349B1 (en) * | 2012-10-30 | 2018-12-19 | Sederma | Combination of plant extracts, cosmetic ingredient and composition containing it and topical cosmetic use thereof |
-
2020
- 2020-12-29 CN CN202011592350.3A patent/CN112704150A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1279727A4 (en) * | 2000-04-26 | 2004-09-01 | Bingxin Wu | Transformation and modification of natural drug by intestinal probiotic fermentative culture |
| CN101099536A (en) * | 2006-07-03 | 2008-01-09 | 河南农业大学 | Application of embedded lactobacillus in animal feed and bacteriostatic promoting production |
| EP2914349B1 (en) * | 2012-10-30 | 2018-12-19 | Sederma | Combination of plant extracts, cosmetic ingredient and composition containing it and topical cosmetic use thereof |
| CN106591182A (en) * | 2016-12-08 | 2017-04-26 | 王丽娟 | Microecological preparation with bacillus subtilis fermenting astragalus membranaceus and application thereof |
| CN106880777A (en) * | 2017-02-27 | 2017-06-23 | 武文海 | A kind of Helth-care product with functions of prolonging life and nourishing face and preparation method thereof |
| CN106666247A (en) * | 2017-03-23 | 2017-05-17 | 威海金牌生物科技股份有限公司 | Foraging feed of special compound feed for breeding stichopus japonicus and preparation method of foraging feed |
| CN107319546A (en) * | 2017-08-07 | 2017-11-07 | 黑龙江省兽医科学研究所 | A kind of fermented tcm powder and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| 冯士彬等: "黄芪多糖对湖羊羔羊生长性能、血清指标、消化功能和直肠菌群的影响", 《江苏农业学报》 * |
| 鲍玉林等: "黄芪多糖对舍饲藏羊生长性能和免疫功能的影响", 《科学技术》 * |
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