Clostridium beijerinckii traditional Chinese medicine fermentation preparation as well as preparation method and application thereof

Abstract

The invention discloses a clostridium beijerinckii traditional Chinese medicine fermentation preparation as well as a preparation method and application thereof, and belongs to the technical field of feed additives and biological fermentation traditional Chinese medicines. Clostridium beijerinckii and powder of five traditional Chinese medicines including astragalus membranaceus, liquorice, poria cocos, atractylodes macrocephala koidz and acanthopanax senticosus are used as raw materials, and anaerobic fermentation culture is performed according to the following steps: S1, inoculating a culture medium with clostridium beijerinckii, culturing for 12-24 hours under an anaerobic condition, then inoculating a culture solution into a bottle according to the proportion of 0.1-0.5%, and culturing for 12-24 hours under the anaerobic condition; carrying out anaerobic culture at 35-40 DEG C for 16-20 hours to obtain a clostridium beijerinckii seed solution; s2, inoculating the clostridium beijerinckii seed liquid into a fermentation tank according to the proportion of 1-5%, and fermenting for 15-20 hours to obtain clostridium beijerinckii fermentation liquid; s3, adding astragalus membranaceus, liquorice, poria cocos, atractylodes macrocephala koidz and acanthopanax senticosus into the clostridium beijerinckii fermentation liquor, and then continuously fermenting and culturing for 10-15 hours to obtain the clostridium beijerinckii traditional Chinese medicine fermentation liquor. The clostridium beijerinckii traditional Chinese medicine fermentation liquor prepared by the invention can effectively improve the growth performance of livestock and poultry, improve the immunity of the livestock and poultry and enhance the disease resistance of the livestock and poultry.

Description

Technical Field

[0001] The invention belongs to the technical field of feed additives and bio-fermented traditional Chinese medicines, and in particular relates to a Clostridium beijerinckii traditional Chinese medicine fermentation preparation, a preparation method and an application thereof. Background Art

[0002] The use of feed antibiotics has greatly promoted the vigorous development of the livestock industry. However, their long-term and extensive use also poses potential biosafety risks to human health and the ecological environment. With technological and social advancements, feed antibiotics are gradually being phased out due to their numerous irreversible drawbacks. The livestock industry is seeking alternatives to feed antibiotics. Against this backdrop, the concepts of "antibiotic ban" and "antibiotic alternatives" have emerged.

[0003] Chinese herbal medicine not only promotes animal growth and regulates animal immunity, but also improves the quality of livestock and poultry products, making it a top choice for antibiotic alternatives. Fermented Chinese herbal medicine is a green and ecological feed additive, obtained by fermenting Chinese herbal medicine using intestinal probiotics.

[0004] Probiotic fermentation can break down macromolecules in Chinese herbal medicine ingredients, converting them into small molecules that can be directly utilized by animals, thereby enhancing their effectiveness. Compared to traditional direct feeding of untreated Chinese herbal medicine additives, fermented Chinese herbal medicine can improve the utilization rate of the active ingredients in Chinese herbal medicines and produce new active ingredients, enhancing the efficacy of use and improving the utilization rate of the effective ingredients in Chinese herbal medicines. Chinese patent application publication number CN 119280330A discloses a Chinese herbal medicine preparation fermented using Clostridium butyricum, and Chinese patent application publication number CN 115287202B discloses a method for fermenting Chinese herbal medicine residue using Kluyveromyces marxianus. In the daily fermentation of Chinese herbal medicines, the utilization rate of Chinese herbal medicine fermentation is limited by the limitations of conventional strains such as yeast and Clostridium butyricum and the fermentation process, which is not conducive to the diversified development of screening and storage of Chinese herbal medicine fermentation strains. In view of this, screening Chinese herbal medicine fermentation strains with excellent storage performance is a key issue we currently need to address.

[0005] Clostridium beijerinckii is a Gram-positive anaerobic bacterium with meso-spore-forming, rod-shaped bodies. The colony has white, filamentous protrusions along its edges and a sticky surface. It primarily produces butyric acid and acetic acid, along with smaller amounts of propionic acid, hydrogen, and carbon dioxide. While C. beijerinckii is primarily used in industrial fermentation to produce butanol, its application in livestock and poultry is relatively limited. Fu Yuze et al. (Fu Yuze, Screening and Identification of Butyric Acid-Producing Clostridium beijerinckii and Its Effects on Lamb Intestinal Health, 2023, Chinese Academy of Agricultural Sciences) identified a strain of C. beijerinckii that reduced inflammatory responses in lambs and enhanced intestinal antioxidant capacity and barrier function. However, there are currently no reports on the use of C. beijerinckii in the fermentation of traditional Chinese medicines. Summary of the Invention

[0006] The present invention aims to provide a traditional Chinese medicine fermentation preparation of Clostridium beijerinckii, as well as its preparation method and application. The traditional Chinese medicine fermentation preparation prepared by the combined fermentation of Clostridium beijerinckii and traditional Chinese medicine can significantly increase the spore conversion rate of Clostridium beijerinckii in the fermentation broth, increase the production of butyric acid and acetic acid, the metabolites of Clostridium beijerinckii, effectively improve the growth performance of livestock and poultry, and enhance their disease resistance.

[0007] To achieve the above object, the present invention adopts the following technical solutions:

[0008] In a first aspect, the present invention provides a method for preparing a Clostridium beijerinckii traditional Chinese medicine fermentation broth, comprising the following steps:

[0009] S1. Inoculate Clostridium beijerinckii into a culture medium, culture under anaerobic conditions for 12-24 hours, then inoculate the culture solution into a bottle at a ratio of 1-5‰, and culture anaerobically at 35-40°C for 16-20 hours to obtain a Clostridium beijerinckii seed solution;

[0010] S2, inoculating the Clostridium beijerinckii seed solution into a fermentation tank at a ratio of 1-5%, adding 65-70% of the fermentation tank volume of fermentation medium, and fermenting at a temperature of 37-40°C, a fermentation pressure of 0.03-0.05 MPa, and a rotation speed of 100-120 r / min for 15-20 hours to obtain Clostridium beijerinckii fermentation liquid;

[0011] S3. Add astragalus, licorice, Poria, Atractylodes macrocephala, and Acanthopanax senticosus to the Clostridium beijerinckii fermentation broth at a mass volume ratio of 2.0-3.0%, wherein the mass ratio of the five Chinese medicinal ingredients, astragalus, licorice, Poria, Atractylodes macrocephala, and Acanthopanax senticosus, is 1-1.5:1-1.5:1-1.5:1-1.5:1-1.5, and then ferment and culture at 35-40°C for 5-15h to obtain the Clostridium beijerinckii fermentation broth.

[0012] Furthermore, in step S1, Clostridium beijerinckii is inoculated into a thioglycollate fluid culture medium and cultured for 18 hours to obtain a culture fluid, wherein the components of the thioglycollate fluid culture medium are: casein peptone 10.0-15.0 g / L, yeast extract powder 3.0-7.0 g / L, glucose 3.0-7.0 g / L, sodium thioglycollate 0.3-0.7 g / L, L-cystine 0.3-0.7 g / L, resazurin 0.0005-0.0015 g / L, and sodium chloride 2.0-3.0 g / L.

[0013] Furthermore, the culture solution was inoculated into a bottle at a ratio of 1‰, and anaerobically cultured at 37°C for 18 hours to obtain a seed solution.

[0014] Furthermore, in step S2, the seed liquid is inoculated into a fermentation tank at a ratio of 2%, and fermented for 18 hours at 37° C., 0.04 MPa, and 110 r / min to obtain a fermentation liquid.

[0015] Furthermore, the fermentation medium components used in the fermentation process in step S2 are: water as the solvent, tryptone 8-10 g / L, yeast extract 2-4 g / L, glucose 15-20 g / L, soluble starch 0.5-1.0 g / L, sodium chloride 2-5 g / L, ammonium citrate 1-2 g / L, Tween-80 0.5-1.0 g / L, dipotassium hydrogen phosphate 0.1-0.3 g / L, potassium dihydrogen phosphate 0.2-0.5 g / L, sodium acetate 1-3 g / L, L-cysteine hydrochloride 0.1-0.5 g / L

[0016] Furthermore, in step S3, astragalus, licorice, poria, atractylodes, and acanthopanax senticosus are added to the fermentation broth at a volume of 2% of the fermentation broth, and then fermented and cultured at 37° C. for 10 h to obtain Clostridium beijerinckii traditional Chinese medicine fermentation broth.

[0017] Furthermore, the mass ratio of the five Chinese medicinal ingredients, namely, astragalus, licorice, poria, atractylodes, and acanthopanax, is 1:1:1:1:1.

[0018] In a second aspect, the present invention further provides a Clostridium beijerinckii traditional Chinese medicine fermentation broth, which is prepared by the above-mentioned preparation method.

[0019] In a third aspect, the present invention further provides the use of the Clostridium beijerinckii traditional Chinese medicine fermentation broth as a livestock and poultry feed additive.

[0020] Furthermore, the amount of Clostridium beijerinckii fermented Chinese medicine liquid added to the livestock and poultry feed additive is 3-5%.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] The Chinese herbal medicine fermentation preparation prepared by the combined fermentation of Clostridium beijerinckii and Chinese herbal medicines of the present invention can significantly improve the spore conversion rate of Clostridium beijerinckii in the fermentation broth, increase the production of butyric acid and acetic acid, the metabolites of Clostridium beijerinckii, effectively improve the growth performance of livestock and poultry, and enhance the disease resistance of livestock and poultry. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is a comparison chart of butyric acid production and pH value of the Clostridium beijerinckii traditional Chinese medicine fermentation broth obtained in Examples 3-7. DETAILED DESCRIPTION

[0024] To better illustrate the present invention, the following embodiments are listed. Obviously, the embodiments described are only part of the present invention, not all of the embodiments. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without making any creative efforts are also within the scope of protection of the present invention.

[0025] The technical solution of the present invention will be further described below with reference to the accompanying drawings and embodiments.

[0026] The experimental methods in the following examples are conventional methods unless otherwise specified. The raw materials involved in the following examples are common commercial products and can be purchased from the market unless otherwise specified.

[0027] The above-mentioned technical features of the present invention and the technical features specifically described below (such as implementation cases) can be combined with each other to form new or preferred technical solutions.

[0028] In the following preparation examples and examples, Clostridium beijerinckii was purchased from the China Industrial Microbiological Culture Collection Center with the number CICC8015.

[0029] Example 1 Clostridium beijerinckii seed culture in a bottle

[0030] Clostridium beijerinckii was stored in glycerol tubes at a concentration of 1.0 × 10 8 CFU / mL. The Clostridium beijerinckii culture medium stored in the glycerol tube was inoculated into 100 mL of thioglycollate fluid culture medium at a ratio of 1%, and then placed in an anaerobic incubator at 37°C for 18 hours. The obtained Clostridium beijerinckii culture medium was inoculated into a 1L triangular flask at a ratio of 1‰, and the triangular flask was filled with thioglycollate fluid culture medium. The triangular flask had a liquid volume of 500 mL and was placed in an anaerobic incubator at 37°C for 18 hours. The final bacterial concentration in the culture medium was 1.5×10 8 CFU / mL.

[0031] The thioglycollate fluid culture medium comprises 15.0 g / L casein peptone, 5.0 g / L yeast extract powder, 5.0 g / L glucose, 0.5 g / L sodium thioglycollate, 0.5 g / L L-cystine, 0.001 g / L resazurin, and 2.5 g / L sodium chloride. Water is used as the solvent, and the pH value is 7.1-7.3.

[0032] Example 2 High-density fermentation culture in a fermenter

[0033] The shake flask seed liquid prepared above was inoculated into the fermenter at a ratio of 2%. The liquid volume of the fermenter was 65% of the fermenter volume. The fermentation temperature was 37°C, the fermentation speed was 110 r / min, the fermenter pressure was 0.04 MPa, and the fermentation was carried out for 18 hours under nitrogen conditions.

[0034] The fermentation tank is filled with a fermentation medium containing the following components: water as a solvent, 9 g / L tryptone, 3 g / L yeast extract powder, 17 g / L glucose, 0.7 g / L soluble starch (acid-hydrolyzed corn soluble starch), 3 g / L sodium chloride, 1.5 g / L ammonium citrate, 0.7 g / L Tween-80, 0.2 g / L dipotassium hydrogen phosphate, 0.3 g / L potassium dihydrogen phosphate, 2 g / L sodium acetate, and 0.3 g / L L-cysteine hydrochloride. At the end of this stage, the spore formation rate of Clostridium beijerinckii is approximately 80-90%.

[0035] Example 3 Preparation of Clostridium beijerinckii Traditional Chinese Medicine Fermentation Broth

[0036] Five kinds of Chinese medicinal powders (directly grinding the Chinese medicinal materials) were added to the Clostridium beijerinckii fermentation broth obtained in Example 2 at a mass volume ratio of 2% g / L. The mass ratio of each component (Astragalus: Licorice: Poria: Atractylodes: Acanthopanax) was 1:1:1:1:1. The culture was continued at 37°C for 10 h to terminate the fermentation. The fermentation broth obtained at this time was the Clostridium beijerinckii Chinese medicinal fermentation broth. The number of viable bacteria in the fermentation broth was 9.2×10 8 CFU / mL.

[0037] Example 4 Preparation of Clostridium beijerinckii Traditional Chinese Medicine Fermentation Broth

[0038] Five kinds of Chinese medicinal powders were added to the Clostridium beijerinckii fermentation broth obtained in Example 2 at a ratio of 2% (same as in Example 3) of the fermentation broth volume, with the ratio of each component (Astragalus: Licorice: Poria: Atractylodes: Acanthopanax) being 1:1:1:1:1. The culture was continued for 15 h to terminate the fermentation. The fermentation broth obtained at this time was the Clostridium beijerinckii Chinese medicinal fermentation broth. The number of viable bacteria in the fermentation broth was 1.17 × 10 9 CFU / mL.

[0039] Example 5 Preparation of Clostridium beijerinckii Traditional Chinese Medicine Fermentation Broth

[0040] Four kinds of Chinese medicinal powders were added to the Clostridium beijerinckii fermentation broth obtained in Example 2 at a ratio of 2% (same as in Example 3) of the fermentation broth volume, with the ratio of each component (Astragalus: Licorice: Atractylodes: Acanthopanax) being 1:1:1:1. The culture was continued at 37°C for 10 h to terminate the fermentation. The fermentation broth obtained at this time was the Clostridium beijerinckii Chinese medicinal fermentation broth. The number of viable bacteria in the fermentation broth was 1.02×10 9 CFU / mL.

[0041] Example 6 Preparation of Clostridium beijerinckii Traditional Chinese Medicine Fermentation Broth

[0042] Five kinds of Chinese medicinal powders were added to the Clostridium beijerinckii fermentation broth obtained in Example 2 at a ratio of 2% (same as in Example 3) of the fermentation broth volume, with the ratio of each component (Astragalus: Licorice: Poria: Atractylodes: Acanthopanax) being 1.5:1:1:1:1. The culture was continued at 37°C for 10 hours to terminate the fermentation. The fermentation broth obtained at this time was the Clostridium beijerinckii Chinese medicinal fermentation broth. The number of viable bacteria in the fermentation broth was 1.04×10 9 CFU / mL.

[0043] Example 7 Preparation of Clostridium beijerinckii Traditional Chinese Medicine Fermentation Broth

[0044] Five kinds of Chinese medicinal powders were added to the Clostridium beijerinckii fermentation broth obtained in Example 2 at a ratio of 2% (same as in Example 3) of the fermentation broth volume, with the ratio of each component (Astragalus: Licorice: Poria: Atractylodes: Acanthopanax) being 1:1.2:1:1:1. The culture was continued at 37°C for 10 h to terminate the fermentation. The fermentation broth obtained at this time was the Clostridium beijerinckii Chinese medicinal fermentation broth. The number of viable bacteria in the fermentation broth was 1.00×10 9 CFU / mL.

[0045] Example 8 Acid Production and pH Detection

[0046] The butyric acid content of the Clostridium beijerinckii traditional Chinese medicine fermentation broth prepared in Example 3-7 was detected using a Huapu high performance liquid chromatograph S3000. The chromatographic conditions were as follows: chromatographic column C18; mobile phase: acetonitrile and 0.1% phosphoric acid solution (injection volume ratio 20:80); injection volume 5 μl; detection wavelength 206 nm; maximum pressure 20 MPa; flow rate 1 ml / min; pH value was detected at the same time. The results are shown in FIG. Figure 1 .

[0047] The results showed that the Clostridium beijerinckii traditional Chinese medicine fermentation broth prepared in Example 3 had the highest butyric acid content and the lowest pH value, which were 13.14 g / L and 4.94, respectively. Therefore, Example 3 was the optimal fermentation condition, that is, the ratio of the five traditional Chinese medicine ingredients (astragalus: licorice: poria: atractylodes: acanthopanax) was 1:1:1:1:1, and the fermentation culture time was 10 h.

[0048] Example 9 Livestock and Poultry Feeding Test

[0049] In the livestock and poultry feeding experiment, the Clostridium beijerinckii traditional Chinese medicine fermentation liquid prepared in Example 3 was used for feeding.

[0050] 1. Piglet feeding test

[0051] Two hundred and fifty healthy, similarly sized, weaned "Du Changda" three-way hybrid piglets (weaned at (21 ± 3) days of age and with an initial weight of (6.50 ± 0.45) kg) were selected and divided into five groups, each containing five replicates (pens) of 10 pigs (5 males and 5 females) with the principle of equal sex and similar weight within each replicate. A blank control group was fed feed alone (without antibiotics); an antibiotic control group was fed feed supplemented with 20 mg / kg colistin sulfate, 50 mg / kg bacitracin zinc, and 75 mg / kg chlortetracycline. The experimental groups were fed a 5% (mL / g) traditional Chinese medicine fermentation preparation mixed with their feed. During the three-day pre-trial period, the experimental pigs were fed the corresponding experimental diet. At the end of the pre-trial period, the pigs were weighed individually and entered the full-scale trial period, which lasted 20 days. The experimental feed was commercially available piglet feed. On the first and 20th days of the trial period, the piglets were fasted for 8 hours (water was not allowed) and then individually weighed to calculate the average daily weight gain. During the trial, the feed consumption was counted in units of replicates (pens), the average daily feed intake, the feed-to-weight ratio, the diarrhea rate and the feeding cost were calculated.

[0052] Table 1 Effects of Chinese herbal fermentation agents on growth performance, diarrhea rate and blood immune indexes of weaned piglets

[0053]

[0054] Note: Data in the same industry with different lowercase letters indicate significant differences (P < 0.05), while data with the same lowercase letters or no letters indicate no significant differences (P > 0.05).

[0055] As shown in Table 1, the experimental group had significantly higher average daily weight gain and feed utilization, and significantly lower diarrhea rates, compared to the blank and antibiotic controls. Blood immunoglobulin IgA and IgG levels were significantly higher in the experimental group than in the blank and antibiotic controls. Therefore, the Clostridium beijerinckii traditional Chinese medicine fermentation culture effectively boosted piglet immunity and reduced diarrhea rates.

[0056] 2. Fattening cattle feeding experiment

[0057] Forty healthy Simmental bulls weighing approximately 600 kg were selected and divided into four groups of 10 each, based on equal weight, fatness, and coat color. A blank control group was fed feed alone, a Clostridium butyricum control group was fed feed supplemented with 5% (mL / g) Clostridium butyricum fermentation broth, and the experimental groups were fed a 5% (mL / g) supplement of a traditional Chinese medicine fermentation preparation. During the 7-day pre-trial period, the experimental fattening cattle were fed the corresponding experimental diet (commercially available bull feed). At the end of the pre-trial period, each animal was weighed and entered the main trial period, which lasted 60 days. In the mornings at the beginning and end of the trial, the cattle were weighed on an empty stomach, and initial and final weights were recorded to calculate average daily gain. Feed intake and feed residue were recorded every 15 days for each group to calculate dry matter intake. Average daily gain and dry matter intake were used to calculate feed-to-weight ratio.

[0058] Table 2 Effects of Chinese herbal fermentation agents on growth performance of fattening cattle

[0059]

[0060] Note: Data in the same industry with different lowercase letters indicate significant differences (P < 0.05), while data with the same lowercase letters or no letters indicate no significant differences (P > 0.05).

[0061] From the results in Table 2, it can be seen that the average daily weight gain of fattening cattle in the experimental group was significantly higher than that in the blank control group and the Clostridium butyricum control group, and the feed-to-weight ratio was significantly lower than that in the blank control group and the Clostridium butyricum control group.

[0062] 3. Meat duck feeding experiment

[0063] One hundred and eighty healthy, one-day-old male Peking ducks of similar body weight were randomly divided into five groups, each containing six replicates. A blank control group was fed a basal diet, a Clostridium butyricum control group was fed a diet supplemented with 5% (mL / g) Clostridium butyricum fermentation broth, and the experimental group was fed a 3% (mL / g) traditional Chinese medicine fermentation preparation. The experimental period lasted 42 days. On days 1, 21, and 42 of feeding, the ducks were fasted for 12 hours and weighed in each replicate. Daily feed consumption was recorded throughout the feeding period. Average daily feed intake, average daily weight gain, feed-to-gain ratio, and serum immunoglobulins (IgA, IgM, and IgG) were calculated for the early (1-21 days), late (21-42 days), and full (1-42 days) periods of the experiment.

[0064] Table 3 Effects of Chinese herbal fermentation agents on growth performance, blood immune indicators, survival rate and disease resistance of meat ducks

[0065]

[0066]

[0067] Note: Data in the same industry with different lowercase letters indicate significant differences (P < 0.05), while data with the same lowercase letters or no letters indicate no significant differences (P > 0.05).

[0068] As shown in Table 3, the daily weight gain of the experimental ducks from 1 to 21 days, 21 to 42 days, and 1 to 42 days was significantly higher than that of the blank control group and the Clostridium butyricum group, and the feed-to-weight ratio was significantly lower than that of the blank control group and the Clostridium butyricum group. Serum immunoglobulin IgA levels in the experimental ducks were significantly higher than those in the blank control group and the Clostridium butyricum group at 30 days. Simultaneously, immunoglobulin IgM and IgG levels in the experimental ducks were slightly higher than those in the blank control group and the Clostridium butyricum group, but the differences were not significant. Furthermore, the diarrhea rate in the experimental ducks was significantly lower than that in the blank control group and the Clostridium butyricum group, and the survival rate was significantly higher than that in the blank control group and the Clostridium butyricum group. Therefore, the Clostridium beijerinckii herbal fermentation culture can improve the immunity of ducks, enhance their resistance to diarrhea, and increase their survival rate.

[0069] The embodiments described above are merely descriptions of preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by persons skilled in the art should fall within the scope of protection defined by the claims of the present invention.

Claims

1. A method for preparing a Clostridium beijerinckii traditional Chinese medicine fermentation broth, characterized in that: The following steps are involved: S1. Inoculate Clostridium beijerinckii into a culture medium, culture under anaerobic conditions for 12-24 hours, then inoculate the culture solution into a bottle at a ratio of 1-5‰, and culture anaerobically at 35-40°C for 16-20 hours to obtain a Clostridium beijerinckii seed solution; S2, inoculating the Clostridium beijerinckii seed solution into a fermentation tank at a ratio of 1-5%, adding 65-70% of the fermentation tank volume of fermentation medium, and fermenting at a temperature of 37-40°C, a fermentation pressure of 0.03-0.05 MPa, and a rotation speed of 100-120 r / min for 15-20 hours to obtain Clostridium beijerinckii fermentation liquid; S3. Add astragalus, licorice, Poria, Atractylodes macrocephala, and Acanthopanax senticosus to the Clostridium beijerinckii fermentation broth at a mass volume ratio of 2.0-3.0%, wherein the mass ratio of the five Chinese medicinal ingredients, astragalus, licorice, Poria, Atractylodes macrocephala, and Acanthopanax senticosus, is 1-1.5:1-1.5:1-1.5:1-1.5:1-1.5, and then ferment and culture at 35-40°C for 5-15h to obtain the Clostridium beijerinckii fermentation broth.

2. The preparation method according to claim 1, characterized in that In step S1, Clostridium beijerinckii is inoculated into a thioglycollate fluid culture medium and cultured for 18 hours to obtain a culture solution. The thioglycollate fluid culture medium comprises: casein peptone 10.0-15.0 g / L, yeast extract powder 3.0-7.0 g / L, glucose 3.0-7.0 g / L, sodium thioglycollate 0.3-0.7 g / L, L-cystine 0.3-0.7 g / L, resazurin 0.0005-0.0015 g / L, and sodium chloride 2.0-3.0 g / L.

3. The preparation method according to claim 2, characterized in that The culture solution was inoculated into a bottle at a ratio of 1‰, and anaerobically cultured at 37°C for 18 hours to obtain a seed solution.

4. The preparation method according to claim 1, characterized in that In step S2, the seed liquid is inoculated into a fermentation tank at a ratio of 2%, and fermented for 18 hours at 37° C., 0.04 MPa, and 110 r / min to obtain a fermentation liquid.

5. The preparation method according to claim 1, characterized in that The fermentation medium used in the fermentation process in step S2 comprises: water as a solvent, 8-10 g / L of tryptone, 2-4 g / L of yeast extract powder, 15-20 g / L of glucose, 0.5-1.0 g / L of soluble starch, 2-5 g / L of sodium chloride, 1-2 g / L of ammonium citrate, 0.5-1.0 g / L of Tween-80, 0.1-0.3 g / L of dipotassium hydrogen phosphate, 0.2-0.5 g / L of potassium dihydrogen phosphate, 1-3 g / L of sodium acetate, and 0.1-0.5 g / L of L-cysteine hydrochloride.

6. The preparation method according to claim 1, characterized in that In step S3, astragalus, licorice, poria, atractylodes, and acanthopanax senticosus are added to the fermentation broth at a volume of 2% of the fermentation broth, and then fermented and cultured at 37° C. for 10 hours to obtain Clostridium beijerinckii traditional Chinese medicine fermentation broth.

7. The preparation method according to claim 6, characterized in that The mass ratio of the five Chinese medicinal ingredients, astragalus, liquorice, poria, atractylodes and acanthopanax, is 1:1:1:1:

1.

8. A Clostridium beijerinckii traditional Chinese medicine fermentation broth, characterized in that: The product is prepared by the preparation method according to any one of claims 1 to 7.

9. Use of the Clostridium beijerinckii traditional Chinese medicine fermentation liquid according to claim 8 as a livestock and poultry feed additive.

10. The use according to claim 9, characterized in that The amount of the Clostridium beijerinckii traditional Chinese medicine fermentation liquid added to the livestock and poultry feed additive is 3-5%.

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