Chinese herbal medicine feed additive for improving production performance of cattle and sheep and preparation method of Chinese herbal medicine feed additive

Abstract

The invention relates to the technical field of feed additives, and particularly discloses a Chinese herbal medicine feed additive for improving the production performance of cattle and sheep and a preparation method thereof. The preparation method comprises the steps of raw material preparation, raw material drying, crushing and screening, raw material mixing, ultrasonic treatment, enzymolysis treatment and fermentation treatment, compared with a traditional method, release of effective bioactive substances in Chinese herbal medicine can be improved, metabolites can be increased after enzymolysis and fermentation treatment, and the effects of resisting inflammation, resisting oxidation and the like are achieved; meanwhile, the Chinese herbal medicine feed additive disclosed by the invention is rich in active ingredients such as flavone, polyphenol and polysaccharide, so that the feed intake of beef cattle and mutton sheep can be increased, the meat-feed ratio can be increased, and the meat growth performance of the productive animals can be further improved.

Description

Technical Field:

[0001] The present invention relates to the technical field of feed additives, and particularly relates to a Chinese herbal medicine feed additive for improving the production performance of cattle and sheep and a preparation method thereof. Background Art:

[0002] Finding green and safe substitutes for growth-promoting functional additives has become a research hotspot in feed additives in recent years.

[0003] Chinese herbal medicine additives refer to those made from plant Chinese herbal medicines as raw materials through different processing methods and having various biological effects. Because Chinese herbal medicine additives have the characteristics of natural sources, diverse functions, safe and reliable use, economy and environmental protection, and compared with traditional additives such as monensin, oxytetracycline, and chlortetracycline, they not only have less toxic effects and no residues, but also can significantly increase the feed intake of animals, improve the production performance of livestock and poultry, improve the feed conversion rate, improve the antioxidant performance and body immunity, and significantly improve the quality of livestock products, etc., and have received extensive attention in the livestock industry.

[0004] Chinese herbal medicines contain various bioactive components, including flavonoids, polysaccharides, organic acids, saponins, alkaloids, etc. The abilities to inhibit bacteria and inflammation, antioxidant, antiviral, enhance body immunity and reduce body stress, etc. are mostly due to the active components. By processing Chinese herbal medicines, the active components of Chinese herbal medicines can be released more fully or produce more metabolites with stronger activity. Traditional processing methods of Chinese herbal medicines such as maceration method, decoction method, percolation method, reflux extraction method, etc., due to poor selectivity, long time, wide extraction range, easy leaching of a large amount of impurities and easy decomposition of active ingredients, directly affect the quality, production efficiency and economic benefits of drugs. Therefore, traditional processing methods can no longer meet the needs of modern production. Summary of the Invention:

[0005] The purpose of the present invention is to provide a Chinese herbal medicine feed additive for improving the production performance of cattle and sheep and a preparation method thereof, so as to improve the release of bioactive components in the Chinese herbal medicine feed additive, reduce impurities, and reduce the decomposition rate of active ingredients.

[0006] The present invention is implemented by the following technical solutions: A preparation method of a Chinese herbal medicine feed additive for improving the production performance of cattle and sheep, comprising the following steps:

[0007] S1: Drying and pulverizing raw materials: Prepare hawthorn, astragalus membranaceus, atractylodes macrocephala, codonopsis pilosula, poria cocos, Chinese yam, euryale ferox, tangerine peel, lotus seed, licorice as raw materials, respectively perform drying treatment on each raw material, and then perform pulverizing treatment on each dried raw material. After pulverizing into fine powder, sieve through a 40-mesh sieve to obtain the raw material powder of each raw material;

[0008] S2: Raw material mixing: Weigh the raw material powders of each raw material in step S1 respectively and mix them to obtain a mixed raw material powder; the raw material powders of each raw material are as follows by weight fraction: 2 - 4 parts of hawthorn, 2 - 4 parts of astragalus membranaceus, 1 - 3 parts of atractylodes macrocephala, 1 - 3 parts of codonopsis pilosula, 1 - 3 parts of poria cocos, 1 - 3 parts of yam, 1 - 3 parts of euryale ferox, 1 - 2 parts of tangerine peel, 1 - 2 parts of lotus seed, 1 - 2 parts of licorice root.

[0009] S3: Ultrasonic treatment: Mix the raw material powder after mixing in step S2 with distilled water, and transfer the mixed slurry to an ultrasonic cell disruptor for ultrasonic treatment to obtain an ultrasonically treated slurry;

[0010] S4: Enzymatic hydrolysis treatment: Add a complex enzyme to the ultrasonically treated slurry obtained in step S3 for enzymatic hydrolysis to obtain an enzymatically hydrolyzed mixture;

[0011] S5: Fermentation treatment: Dehydrate and dry the enzymatically hydrolyzed mixture obtained in step S4 to obtain an enzymatically hydrolyzed dry powder, place the enzymatically hydrolyzed dry powder in a fermentation container, add water and mix evenly, then add a complex bacterial liquid for fermentation, and obtain a Chinese herbal medicine feed additive after fermentation is completed.

[0012] Further, the drying condition in step S1 is drying at 60°C - 65°C for 4 - 8 hours.

[0013] Further, in step S3, the raw material powder and distilled water are mixed at a liquid - solid ratio of 10∶1.

[0014] Further, the conditions for ultrasonic treatment of the ultrasonic cell disruptor in step S3 are ultrasonic power of 400 - 600 W and ultrasonic time of 25 - 45 min.

[0015] Further, the complex enzyme in step S4 is a complex enzyme preparation composed of pectinase > 30 U / g, xylanase > 1200 U / g, β - glucanase > 200 U / g, and cellulase > 30 U / g.

[0016] Further, the added mass of the complex enzyme in step S4 is 0.2 - 0.5% of the mass of the mixed raw material powder in step S2, the temperature for enzymatic hydrolysis is 30 - 50°C, and the enzymatic hydrolysis treatment time is 4 - 6 h.

[0017] Further, the mass of water added in step S5 is 20 - 50% of the mass of the enzymatically hydrolyzed dry powder.

[0018] Further, the complex bacterial liquid in step S5 is a complex bacterial liquid of bacillus subtilis and bacillus licheniformis.

[0019] Further, the fermentation conditions in step S5 are: the added mass of the complex bacterial liquid is 3 - 5% of the mass of the enzymatically hydrolyzed dry powder, and the fermentation days are 3 - 5 days.

[0020] A Chinese herbal medicine feed additive for improving the production performance of cattle and sheep, which is prepared by the preparation method of the Chinese herbal medicine feed additive for improving the production performance of cattle and sheep described above.

[0021] Advantages of the present invention:

[0022] (1) The Chinese herbal medicine feed additive processed from raw materials including hawthorn, astragalus membranaceus, atractylodes macrocephala, codonopsis pilosula, poria cocos, Chinese yam, euryale ferox, tangerine peel, lotus seeds, and liquorice is rich in active ingredients such as flavonoids, polyphenols, and polysaccharides, which can increase the feed intake of beef cattle and sheep, improve the meat-to-feed ratio. The Chinese herbal medicine feed additive disclosed in the present invention has the functions of improving animal immunity, antioxidation, and reducing liver damage in animals. Adding it to the feed can promote the digestion of dietary nutrients by animals, promote protein synthesis metabolism in the body, enhance the deposition of fat and nitrogen, and thus improve the meat growth performance of such productive animals.

[0023] (2) By pulverizing, ultrasonic treatment, enzymatic hydrolysis treatment, and fermentation treatment of Chinese herbal medicine raw materials, the active ingredients of Chinese herbal medicine can be released more fully. More metabolites with stronger activity are produced during the fermentation process. Compared with the traditional method of processing Chinese herbal medicine raw materials, more bioactive ingredients can be retained, the dosage of Chinese herbal medicine feed additive can be reduced, and economic benefits can be improved. Description of the drawings:

[0024] Figure 1 It is the PCA score chart of the Chinese herbal medicine feed additive prepared in Example 1 and Comparative Example 4 of the present invention.

[0025] Figure 2 It is the clustering heat map of the Chinese herbal medicine feed additive prepared in Example 1 and Comparative Example 4 of the present invention.

[0026] Figure 3 It is the dynamic distribution chart of the difference in metabolite content of the Chinese herbal medicine feed additive prepared in Example 1 and Comparative Example 4 of the present invention.

[0027] Figure 4 It is the schematic diagram of the DPPH scavenging rate of the Chinese herbal medicine feed additive prepared in Example 1 of the present invention. Detailed implementation manners:

[0028] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments; based on the embodiments in the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative efforts belong to the scope of protection of the present invention.

[0029] Unless otherwise specified, the raw materials used in the embodiments of the present invention are commercially available Chinese herbal medicine raw materials, and reagents and the like are ordinary commercially available products. The following is only an exemplary illustration:

[0030] The complex enzyme is purchased from Boyide Biotechnology Co., Ltd.; the complex bacterial liquid is purchased from Boyide Biotechnology Co., Ltd.

[0031] Example 1: A Chinese herbal medicine feed additive for improving the production performance of cattle and sheep. The specific preparation steps are as follows:

[0032] S1: Drying and pulverizing the raw materials: Prepare hawthorn, astragalus membranaceus, atractylodes macrocephala, codonopsis pilosula, poria cocos, Chinese yam, euryale ferox, tangerine peel, lotus seeds, and licorice as raw materials. Each raw material is dried, and each dried raw material is pulverized into fine powder and passed through a 40-mesh sieve to obtain the raw material powder of each raw material. The raw material drying conditions are: drying at 65°C for 4 hours;

[0033] S2: Mixing the raw materials: Take the sieved raw material powder obtained in S1 above, and mix it according to the following mass ratio: 2 parts of hawthorn, 2 parts of astragalus membranaceus, 1 part of atractylodes macrocephala, 1 part of codonopsis pilosula, 1 part of poria cocos, 1 part of Chinese yam, 1 part of euryale ferox, 1 part of tangerine peel, 1 part of lotus seeds, and 1 part of licorice to obtain the mixed raw material powder;

[0034] S3: Ultrasonic treatment: Mix the mixed raw material powder obtained in S2 above with distilled water according to a liquid-solid ratio of 10:1, and place it in an ultrasonic cell disruptor with an ultrasonic power of 400 W and an ultrasonic time of 45 min to obtain an ultrasonic-treated slurry; (The ultrasonic cell disruptor is manufactured by Ningbo Xinzhi Biotechnology Co., Ltd., model JY92-IIDN)

[0035] S4: Add a complex enzyme accounting for 0.3% of the mass of the mixed raw material powder in the ultrasonic-treated slurry obtained in S3 above. The specific components of the complex enzyme are: pectinase > 30 U / g, xylanase > 1200 U / g, β-glucanase > 200 U / g, cellulase > 30 U / g. The complex enzyme is purchased from Boyide Biotechnology Co., Ltd., and the ultrasonic-treated slurry is subjected to enzymatic hydrolysis treatment. The enzymatic hydrolysis temperature is controlled at 45°C and the enzymatic hydrolysis is carried out for 6 hours to obtain an enzymatically hydrolyzed mixture;

[0036] S5: Dehydrate and dry the enzymatically hydrolyzed mixture obtained in step S4 to obtain an enzymatically hydrolyzed dry powder. Place the enzymatically hydrolyzed dry powder in a fermentation container, add water accounting for 40% of the mass of the enzymatically hydrolyzed dry powder and mix evenly, and then add a complex bacterial liquid accounting for 4% of the mass of the enzymatically hydrolyzed dry powder for fermentation. The type of complex bacterial liquid added is bacillus subtilis and bacillus licheniformis. The complex bacterial liquid is purchased from Boyide Biotechnology Co., Ltd., and the fermentation days are 4 days. After the fermentation is completed, the Chinese herbal medicine feed additive is obtained.

[0037] Example 2: A Chinese herbal medicine feed additive for improving the production performance of cattle and sheep. The difference in the specific preparation steps from Example 1 lies in that in step S2, the mass fraction ratios of various raw material powders are different. Among them, there are 4 parts of hawthorn, 4 parts of astragalus membranaceus, 3 parts of atractylodes macrocephala, 3 parts of codonopsis pilosula, 3 parts of poria cocos, 3 parts of Chinese yam, 3 parts of euryale ferox, 2 parts of dried tangerine peel, 2 parts of lotus seeds, and 2 parts of licorice; the remaining operation steps are the same as those in Example 1.

[0038] Example 3: A Chinese herbal medicine feed additive for improving the production performance of cattle and sheep. The difference in the specific preparation steps from Example 1 lies in that in step S2, the mass fraction ratios of various raw material powders are different. Among them, there are 3 parts of hawthorn, 3 parts of astragalus membranaceus, 2 parts of atractylodes macrocephala, 2 parts of codonopsis pilosula, 3 parts of poria cocos, 2 parts of Chinese yam, 2 parts of euryale ferox, 2 parts of dried tangerine peel, 1 part of lotus seeds, and 1 part of licorice.

[0039] Example 4: A Chinese herbal medicine feed additive for improving the production performance of cattle and sheep. The difference in the specific preparation steps from Example 1 lies in that in step S3, the ultrasonic power in the ultrasonic cell disruptor for ultrasonic treatment is 600 W, and the ultrasonic time is 45 min. The remaining operation steps are the same as those in Example 1.

[0040] Example 5: A Chinese herbal medicine feed additive for improving the production performance of cattle and sheep. The difference in the specific preparation steps from Example 1 lies in that in step S3, the ultrasonic power in the ultrasonic cell disruptor for ultrasonic treatment is 400 W, and the ultrasonic time is 25 min. The remaining operation steps are the same as those in Example 1.

[0041] Comparative Example 1: A Chinese herbal medicine feed additive for improving the production performance of cattle and sheep. The raw materials selected are the same as those in Example 1. The difference in the specific preparation steps from Example 1 lies in that no enzymatic hydrolysis and fermentation treatment are carried out. Its steps S1 - S3 are the same as those in Example 1, and the ultrasonic treatment slurry obtained in step S3 is the Chinese herbal medicine feed additive prepared in this comparative example.

[0042] Comparative Example 2: A Chinese herbal medicine feed additive for improving the production performance of cattle and sheep. The raw materials selected are the same as those in Example 1. The difference in the specific preparation steps from Example 1 lies in that no ultrasonic and enzymatic hydrolysis treatment with composite enzymes is carried out. Steps S1 - S2 are the same as those in Example 1. In step S3, the mixed raw material powder obtained in step S2 is directly placed into a fermentation container, 40% of the water by the mass of the mixed raw material powder is added and mixed evenly, and then composite bacterial liquid is added for fermentation. The types of composite bacterial liquid added are bacillus subtilis and bacillus licheniformis, purchased from Boyide Biotechnology Co., Ltd. The addition amount of the composite bacterial liquid is 4% of the mass of the mixed raw material powder, and the fermentation days are 4 days. After the fermentation is completed, the Chinese herbal medicine feed additive of this comparative example is obtained.

[0043] Comparative Example 3: A Chinese herbal medicine feed additive for improving the production performance of cattle and sheep. The raw materials selected are the same as those in Example 1. The specific preparation steps are different from those in Example 1 in that ultrasonic and fermentation treatments are not carried out. Steps S1 - S2 are the same as those in Example 1. In step S3, the mixed raw material powder obtained in step S2 is mixed with distilled water according to a liquid - solid ratio of 10∶1 and placed in a container to obtain a mixed slurry. Then, a compound enzyme accounting for 0.3% of the mass of the mixed raw material powder in S2 is added. The specific components of the compound enzyme are: pectinase > 30U / g, xylanase > 1200U / g, β - glucanase > 200U / g, cellulase > 30U / g, purchased from Boyide Biotechnology Co., Ltd. The slurry treated by ultrasonic is subjected to enzymatic hydrolysis treatment. The enzymatic hydrolysis temperature is controlled at 45°C and the enzymatic hydrolysis is carried out for 6 hours to obtain an enzymolysis mixture. The obtained enzymolysis mixture is the Chinese herbal medicine feed additive of this comparative example.

[0044] Comparative Example 4: A Chinese herbal medicine feed additive for improving the production performance of cattle and sheep. The raw materials selected are the same as those in Example 1. The specific preparation steps are different from those in Example 1 in that fermentation treatment is not carried out. Steps S1 - S4 are the same as those in Example 1. The enzymolysis mixture obtained in step S4 is the Chinese herbal medicine feed additive of this comparative example.

[0045] Comparative Example 5: A Chinese herbal medicine feed additive prepared by traditional water extraction method. The raw materials selected are the same as those in Example 1. It specifically includes the following steps: S1: Commercially available hawthorn, astragalus root, atractylodes rhizome, codonopsis pilosula, poria cocos, Chinese yam, euryale ferox, tangerine peel, lotus seed, licorice root are used as raw materials and are respectively dried. Drying conditions: drying at 65°C for 4 hours. Each dried raw material is respectively pulverized and ground into fine powder and then sieved through a 40 - mesh sieve to obtain the raw material powder of each raw material; S2: Take the sieved raw material powder obtained in S1 above and mix them according to the following mass ratio: 2 parts of hawthorn, 2 parts of astragalus root, 1 part of atractylodes rhizome, 1 part of codonopsis pilosula, 1 part of poria cocos, 1 part of Chinese yam, 1 part of euryale ferox, 1 part of tangerine peel, 1 part of lotus seed, 1 part of licorice root to obtain a mixed raw material powder; Add water and stir to mix to obtain a mixed slurry, where the liquid - solid ratio of water to the mixed raw material powder is 10∶1; Then, the mixed slurry is heated for water extraction. The water extraction time is 30 min and the water extraction temperature is 90°C. The water extract obtained is the Chinese herbal medicine feed additive of this comparative example.

[0046] To illustrate the effective bioactive substances in the Chinese herbal medicine feed additive for improving the production performance of cattle and sheep in the embodiments of the present invention, in vitro effective component experiments were carried out on the feed additives prepared in the above - mentioned embodiments and comparative examples. The specific experimental methods are as follows:

[0047] Weigh 10 g of the Chinese herbal medicine feed additives prepared in Examples 1 - 5 and Comparative Examples 1 - 3 of the present invention into conical flasks, and respectively detect the flavonoid, polyphenol, and polysaccharide contents. The specific experimental methods are as follows:

[0048] Determination of flavonoid content

[0049] Respectively pipette 0.1 mL of the supernatant of the Chinese herbal medicine feed additives of Examples 1-5 and Comparative Examples 1-5 after centrifugation, and place them in 25-mL volumetric flasks respectively. The aluminum salt complexation reaction is adopted: add 1 mL of 5% sodium nitrite solution, mix well, and let stand for 6 min. Then add 1 mL of 10% aluminum nitrate, mix well, and let stand for 6 min. Finally, add 10 mL of 10% sodium hydroxide, make up the volume to the scale with ultrapure water, let stand for 15 min, and measure the absorbance at a wavelength of 510 nm. The regression equation of the flavonoid standard curve is: y = 1.2725X1 + 0.0044. In the formula, y corresponds to the absorbance, and X1 corresponds to the mass concentration. Calculate the flavonoid mass concentration according to the absorbance and the flavonoid standard curve, and calculate the flavonoid content according to the following formula.

[0050] The calculation formula is

[0051] Y is the content of the active ingredient of the compound Chinese herbal medicine feed additive (mg / g); X1 is the mass concentration of flavonoids in the supernatant of the Chinese herbal medicine feed additive taken (mg / mL); V is the volume of the solution (mL); N is the dilution factor; M is the mass of the Chinese herbal medicine feed additive (g).

[0052] (2) Determination of polyphenol content

[0053] Respectively pipette 0.1 mL of the supernatant of the Chinese herbal medicine feed additives of Examples 1-5 and Comparative Examples 1-5 after centrifugation, and place them in 10-mL colorimetric tubes respectively. Add 2.5 mL of Folin-Ciocalteu reagent, shake well, then add 2.5 mL of 15% sodium carbonate solution, titrate to the scale line with distilled water, water bath at 40 °C for 60 min, then cool and let stand for 20 min, and measure the absorbance of the solution at a wavelength of 778 nm. The regression equation of the polyphenol standard curve is: y = 154.55X2 + 0.023. In the formula, y corresponds to the absorbance, and X2 corresponds to the mass concentration. Calculate the polyphenol mass concentration according to the absorbance and the polyphenol standard curve, and calculate the polyphenol content according to the following formula.

[0054] The calculation formula is

[0055] Y is the content of the active ingredient of the compound Chinese herbal medicine feed additive (mg / g); X2 is the mass concentration of polyphenols in the supernatant of the Chinese herbal medicine feed additive taken (mg / mL); V is the volume of the solution (mL); N is the dilution factor; M is the mass of the Chinese herbal medicine feed additive (g)

[0056] (3) Determination of polysaccharide content

[0057] Respectively, 0.1 mL of the supernatant of the Chinese herbal medicine feed additives in Examples 1-5 and the Chinese herbal medicine feed additives in Comparative Examples 1-5 after centrifugation were taken and placed in 10 mL volumetric flasks. Distilled water was added, shaken well and made up to the scale line to obtain the working solution of the Chinese herbal medicine additive. Exactly 0.1 mL of the working solution of the Chinese herbal medicine additive was taken and placed in a 10 mL colorimetric tube. 1 mL of 5% phenol solution was added, shaken well, then 5 mL of concentrated sulfuric acid solution was added, shaken well and made up to the scale line. It was placed in a boiling water bath for 15 min, taken out and cooled to room temperature. The absorbance of the solution was measured at a wavelength of 490 nm. The regression equation of the polysaccharide standard curve was: y = 13.853X3 + 0.0542. In the formula, y corresponds to the absorbance, and X3 corresponds to the mass concentration. The polysaccharide mass concentration was calculated according to the absorbance and the polysaccharide standard curve, and the polysaccharide content was calculated according to the following formula.

[0058] The calculation formula is

[0059] Y is the content of the active ingredient in the compound Chinese herbal medicine feed additive (mg / g); X3 is the mass concentration of polysaccharide in the supernatant of the Chinese herbal medicine feed additive taken (mg / mL); V is the volume of the solution (mL); N is the dilution factor; M is the mass of the Chinese herbal medicine feed additive (g)

[0060] The contents of the effective bioactive substances in the Chinese herbal medicine feed additives prepared in Examples 1-5 and Comparative Examples 1-5 are shown in Table 1

[0061] Table 1

[0062]

[0063]

[0064] By comparing the effective bioactive substances in the Chinese herbal medicine feed additives of Example 1 and Comparative Examples 1-4, the contents of flavonoids, polysaccharides and polyphenols in the Chinese herbal medicine feed additives can be effectively increased after the combined treatment of ultrasonic treatment, enzymatic hydrolysis treatment and fermentation treatment. The specific mechanisms of action of each treatment method on the Chinese herbal medicine raw materials are as follows:

[0065] Ultrasonic treatment uses the energy of ultrasound to physically disrupt the cell wall structure of Chinese herbal medicines, exposing the active ingredients in the Chinese herbal medicines, thereby achieving the effect of improving the efficiency of subsequent enzymatic hydrolysis. The working principle of ultrasonic treatment is to generate high-frequency sound waves, creating tiny bubbles (cavitation) in the liquid. When these bubbles burst, they release a large amount of energy in the form of heat and shock waves. Ultrasonic waves of 400 - 600W provide a strong enough effect to break plant cells and release bioactive compounds such as alkaloids, flavonoids, and essential oil compounds without causing excessive loss of these compounds into the surrounding medium. At the same time, since Chinese herbal medicines usually consist of complex plant structures, including hard cell walls made of cellulose, lignin, and other polysaccharides. It takes time to effectively break the cell wall and release the active ingredients (such as alkaloids, flavonoids, essential oils, and polysaccharides). This process is not instantaneous. If the time is too short, not all particles in the herbal mixture will be fully exposed to the cavitation effect, resulting in insufficient fragmentation. If the time is too long, heat-sensitive compounds in the herbal extract, such as vitamins, enzymes, and antioxidants, will be damaged. Therefore, based on the plant cell wall structure of the raw materials selected in the present invention and related experiments, it is most suitable that the ultrasonic time is 25 - 45 minutes.

[0066] The optimal process for enzymatic hydrolysis of compound Chinese herbal medicines by composite enzymes is that the enzyme addition amount is 0.3%, the enzymatic hydrolysis time is 6 hours, and the enzymatic hydrolysis temperature is 45°C. The enzymatic hydrolysis reaction can break down the plant cell wall of Chinese herbal medicines and accelerate the rapid release of active ingredients (such as polyphenols, flavonoids, volatile oils, etc.) in Chinese herbal medicines. Specifically: Pectinase can split pectin molecules, making the cell wall looser, and the active ingredients in Chinese herbal medicines are more easily released, improving the overall efficacy of Chinese herbal medicines; Cellulase can effectively degrade the cellulose in herbs, releasing polyphenols, flavonoids, and polysaccharides wrapped by cellulose, thereby improving the pharmacological activity of herbs; In addition, when cellulase degrades cellulose, it produces sugars, which can be used as carbon sources during the fermentation process to promote the growth and proliferation of lactic acid bacteria. β - Glucanase can convert it into small molecule oligosaccharides of glucan, which are easier to preserve and be absorbed by the body, thus improving the efficacy of herbs; Xylanase mainly degrades the lignin in the plant cell wall, destroys the structure of plant wall cells, and improves the extractability and bioavailability of active substances in herbs.

[0067] Bacillus subtilis and Bacillus licheniformis metabolize to produce various enzymes, such as cellulase, pectinase, ligninase, and lipase, all of which have high catalytic efficiency and can decompose various complex organic substances in Chinese herbal medicines, including cellulose, starch, protein, etc., resulting in an increase in the synthesis of antioxidant components such as polyphenols and flavonoids in Chinese herbal medicines, enhancing the antioxidant effect and immunomodulatory effect of the herbs. Moreover, the volatile substances and aromatic components produced during the microbial fermentation process can improve the flavor of the herbs, increase palatability, and increase the feed intake of livestock. Bacillus subtilis and Bacillus licheniformis can also convert flavonoids, steroid compounds, etc. into forms that are more easily absorbed and utilized, thereby improving the utilization efficiency of the herbs. In addition, Bacillus subtilis and Bacillus licheniformis in the compound bacterial powder can inhibit the growth of pathogenic bacteria during the fermentation process, prevent contamination and spoilage, and thus improve the safety of the fermentation products.

[0068] By comparing the effective bioactive substances in the Chinese herbal medicine feed additives of Example 1 and Comparative Example 5, compared with the traditional water extraction method, the method of Example 1 of the present invention can greatly increase the content of effective bioactive substances in the Chinese herbal medicine feed additives.

[0069] In order to further illustrate the principle of the release and processing of effective bioactive components in Chinese herbal medicines by fermentation, the active components of the Chinese herbal medicine feed additives prepared in Example 1 and Comparative Example 4 above were detected using gas chromatography-mass spectrometry (GC-MS):

[0070] The Chinese herbal medicine feed additive disclosed in Example 1 of the present invention and the Chinese herbal medicine feed additive of Comparative Example 4 were placed in a freeze dryer (Scientz-100F) for vacuum freeze drying, and ground (30 Hz, 1.5 min) using a grinder (MM400, Retsch) until it became powdery; 100 mg of the powder was weighed and dissolved in 1.2 mL of 70% methanol extract, vortexed once every 30 minutes for 30 seconds each time, and vortexed a total of 6 times. The sample was placed in a 4°C refrigerator overnight; after centrifugation (at a speed of 12,000 rpm for 10 minutes), the supernatant was aspirated, the sample was filtered through a microporous membrane (0.22 μm pore size), and stored in a sample injection bottle for (UPLC-MS / MS) analysis. The obtained PCA score plot is as Figure 1 and the clustering heat map is as Figure 2 shown. In the figure, CG represents Comparative Example 4; EG represents Example 1, and mix01, mix02, and mix03 are quality control samples.

[0071] Analysis of active ingredients in fermented and unfermented Chinese herbal medicines by widely targeted metabolomics technology identified a total of 1,275 metabolites. From the PCA score plot, it can be seen that the data points of Example 1 of the invention and Comparative Example 4 are clearly distinguishable in the figure. By performing cluster analysis on the content values of their metabolites, it can be seen that there are obvious differences between the two, and they are clustered into two categories, indicating that the metabolites of the Chinese herbal medicine feed additive disclosed in Example 1 are significantly different from those of the Chinese herbal medicine feed additive in Comparative Example 4.

[0072] By calculating the fold change of metabolites in the two groups, a dynamic distribution map of metabolite content differences was obtained, as Figure 3 shown.

[0073] Among them, there are 210 up-regulated metabolites and 349 down-regulated metabolites. This shows that there are obvious changes in the metabolites of the Chinese herbal medicine feed additive disclosed in Example 1 of the invention compared with Comparative Example 4; the screening results of some differential metabolites are shown in Table 2:

[0074] Table 2: Screening results of some differential metabolites

[0075]

[0076]

[0077] Among the differential metabolites, there are a total of 32 isoflavonoid compounds, with 16 up-regulated metabolites and 16 down-regulated metabolites each. Among the up-regulated metabolites, pterocarpin has strong antibacterial, antifungal, antioxidant, antitumor and other biological activities; genistein is a kind of phytoestrogen with a molecular structure similar to estrogen, mainly present in the stems, leaves and seeds of leguminous plants such as soybeans and alfalfa. Due to the presence of a polyhydroxyphenol structure in its structure, it has certain antioxidant biological activities. Among the down-regulated metabolites, sophoricoside mainly comes from leguminous plants and has an anti-fertility effect and toxic effect. Among the up-regulated metabolites, kaempferol has good antioxidant and anti-inflammatory abilities. Kaempferol has a significant effect on inhibiting the proliferation of pancreatic cancer cells, can induce apoptosis of cancer cells, and at the same time can make cancer cells sensitive to chemotherapy, indicating that kaempferol may have a clinical role in the treatment of pancreatic cancer. Quercetin exists mostly in the form of glycosides in plants and has antioxidant and anti-inflammatory effects.

[0078] There are a total of 78 phenolic acid compounds among the differential metabolites, including 50 up-regulated metabolites and 28 down-regulated metabolites. Among the up-regulated metabolites, benzoic acid compounds are the simplest aromatic acids with a carboxyl group directly connected to the benzene ring. Gallic acid has excellent antibacterial, antiviral, and anti-tumor effects and can scavenge excessive reactive free radicals in the body. Ferulic acid has excellent antioxidant capacity. Adding free ferulic acid to the lamb diet can improve its meat quality, reduce the protein oxidation index to a certain extent, and improve the feed utilization rate. Chlorogenic acid and its isomers such as cryptochlorogenic acid and isochlorogenic acid have antioxidant capacity because they contain phenolic hydroxyl groups.

[0079] There are a total of 49 alkaloids among the differential metabolites, including 23 up-regulated metabolites and 26 down-regulated metabolites. Among the up-regulated metabolites, sophocarpine is mostly derived from Chinese herbal medicines such as Sophora flavescens and Sophora alopecuroides and has good biological activities such as immunomodulatory and anti-tumor activities. Demethylcoclaurine belongs to the structural class of berberine and has effects such as lowering blood pressure.

[0080] There are a total of 24 carbohydrates among the differential metabolites, including 10 up-regulated differential metabolites and 14 down-regulated metabolites. Among the up-regulated metabolites, rhamnose has good anti-fungal and anti-tumor activities. Glucuronolactone can combine with some toxic substances in the body to form non-toxic glucuronic acid, which can effectively prevent and treat diseases such as liver cirrhosis and has an obvious effect of protecting the liver and detoxifying. Mannotriose is a kind of oligosaccharide, and its use as a feed additive can replace antibiotics to promote the growth of the body.

[0081] The discovery of these metabolites can be used to illustrate the nutritional characteristics and functions of the Chinese herbal medicine feed additive disclosed in Example 1 of the present invention. Compared with the Chinese herbal medicine feed additive in Comparative Example 4, the Chinese herbal medicine feed additive disclosed in Example 1 of the present invention has more active substances and a much higher content than before treatment. It shows that the Chinese herbal medicine feed additive disclosed in Example 1 of the present invention significantly enhances the efficacy and functions of Chinese herbal medicines.

[0082] Free radicals are the main culprits leading to cell membrane oxidation, promoting aging, and DNA damage; the Chinese herbal medicine feed additive disclosed in the present invention has the effect of reducing DPPH free radicals in the body. In order to illustrate this effect, the DPPH free radical scavenging rate of the Chinese herbal medicine feed additive prepared in Example 1 of the present invention was measured, and the specific operation is as follows:

[0083] Accurately measure 2.5 mg of DPPH. DPPH (2,2-Diphenyl-1-picrylhydrazyl) is a stable free radical that can be used to measure the free radical scavenging activity of antioxidants. Dissolve it with methanol and make up the volume to 100 mL in a volumetric flask to obtain a stock solution with a concentration of 25 μg / mL, which should be prepared freshly before use. Respectively take 0, 2, 4, 6, 8, 10 mL of the stock solution and place them in 10 mL volumetric flasks, then make up the volume to the calibration line with methanol to obtain standard solutions with concentrations of 0, 5, 10, 15, 20, 25 μg / mL. Measure the absorbance at a wavelength of 515 nm on an ultraviolet spectrophotometer, plot the standard curve and obtain the regression equation.

[0084] Prepare the Chinese herbal medicine feed additive disclosed in Example 1 of the present invention with concentrations of 0.02, 0.06, 0.10, 0.12, 0.16 mg / mL. Use ascorbic acid as a reference substance and prepare solutions with the same concentrations. Take 0.1 mL of the above solutions into test tubes, then add 3.9 mL of DPPH solution with a concentration of 25 μg / mL, and use methanol solution to replace the test solution as a blank. Measure the absorbance at a wavelength of 515 nm using an ultraviolet spectrophotometer, and then obtain the DPPH concentration according to the DPPH standard curve.

[0085] Calculation formula: DPPH·REM = DPPH·r / DPPH·r0 × 100%

[0086] Where DPPH·r is the mass concentration of DPPH· after adding the antioxidant and stabilizing, and DPPH·r0 is the original mass concentration of DPPH·.

[0087] From Figure 4 it can be seen that when the extract concentration in the group of Example 1 of the invention is in the range of 0.02 - 0.10 mg / g, the DPPH scavenging rate gradually increases, reaches a plateau at 0.12 - 0.16 mg / g, and the highest reaches 50.64% of ascorbic acid; this shows that the Chinese herbal medicine feed additive disclosed in Example 1 of the invention has good antioxidant performance and the function of scavenging free radicals in organisms.

[0088] In order to illustrate that the Chinese herbal medicine feed additive for improving the production performance of beef cattle and sheep in Example 1 of the present invention can improve the growth performance of animals, promote the digestion of dietary nutrients by animals, promote protein synthesis metabolism in the body, enhance the deposition of fat and nitrogen, and at the same time reduce liver damage in animals and improve the immune function of animals, feeding experiments on two animals, beef cattle and mutton sheep, were respectively carried out. The specific experimental methods are as follows:

[0089] Feeding experiment one:

[0090] 1. Experimental animals

[0091] In Feeding Experiment 1, 24 beef cattle aged 24 - 26 months, in good body condition and with a body weight of 383.67 ± 34.06 kg were selected as experimental animals. They were uniformly immunized and dewormed, and raised in single pens. Feeding was carried out twice a day at 8:00 and 18:00, with free access to food and water. The pre-feeding period was 15 days.

[0092] 2. Composition of the basal diet

[0093] The design of the animal basal diet in Feeding Experiment 1 referred to the Feeding Standard for Beef Cattle in China (NY / T 815 - 2004). The composition of the diet and its nutritional level components are shown in Table 3.

[0094] Table 3 Nutritional levels of the diet (dry matter basis)

[0095]

[0096]

[0097] The net energy for maintenance (NE0mf) was a calculated value (referring to the Feeding Standard for Beef Cattle in "NY / T 815 - 2004" and calculating the comprehensive net energy value of raw materials using the "Table of Common Feed Ingredients and Nutritional Values for Beef Cattle"), and the remaining nutritional indicators were all measured values.

[0098] 3. Experimental design

[0099] The beef cattle were randomly divided into 4 groups, with 6 heads in each group. Group I was the blank control group fed with the basal diet, Group II was fed with the feed added with the Chinese herbal medicine feed additive prepared in Comparative Example 5 at 0.2% of the weight of the basal diet, Group III was fed with the feed added with the Chinese herbal medicine feed additive disclosed in Example 1 of the present invention at 0.2% of the weight of the basal diet, and Group IV was fed with the feed added with the untreated Chinese herbal medicine feed additive at 0.2% of the weight of the basal diet (the preparation method of the untreated Chinese herbal medicine feed additive was: selecting the same proportion of Chinese medicine raw materials as in Example 1, grinding and pulverizing them, passing through a 40 - mesh sieve, and directly adding the obtained Chinese medicine raw material powder to the feed); the experimental period was 60 days.

[0100] 4. Sampling of samples

[0101] Collection and treatment of feed raw materials: Feed samples were collected and dried to a constant weight at 65 °C, and then pulverized through a 40 - mesh sieve as analysis samples.

[0102] Fecal sample collection and pretreatment: After the 30th day of the formal experiment, 100 g of cow fecal samples were collected from each cow every morning before feeding, continuously for 10 days. The fecal samples collected every day were divided into two parts. One part was added with 10% H2SO4 for nitrogen fixation for crude protein determination, and the other part was stored frozen at -20 °C. At the end of the experiment, the 10-day fecal samples of each cow were mixed evenly. After taking a part and drying it at 65 °C to determine the initial moisture, it was pulverized with a traditional Chinese medicine pulverizer and passed through a 40-mesh sieve to be used as a fecal sample analysis sample for routine nutrient composition analysis.

[0103] Serum preparation: Before morning feeding at the end of the formal trial period, 5 ml of blood was collected from the jugular vein of the experimental cows in each group, taking care to avoid shaking to prevent hemolysis. Then it was centrifuged at a low speed of 4000 r / min for 10 min, and the serum was transferred and aliquoted into 1.5 ml enzyme-free and sterile centrifuge tubes and stored frozen at -80 °C for later measurement.

[0104] 5. Determination indexes

[0105] (1) Determination of beef cattle growth performance

[0106] After the start of the formal feeding period of the experiment, the feed intake of each group of cows was strictly recorded every week. The cows were weighed on an empty stomach in the morning at the start and end of the experiment, and the experimental data were recorded. Calculate:

[0107] Average daily feed intake (ADFI) = total feed intake / total number of days of the experiment

[0108] Average daily gain (ADG) = total weight gain during the experimental period / number of days of the experiment

[0109] Feed to gain ratio (F / G) = daily feed weight / daily feed intake / daily gain

[0110] The specific measurement results are shown in Table 4 below:

[0111] Table 4

[0112]

[0113] Note: In the same row, without letter or data superscripts, the same letter indicates no significant difference (P > 0.05), different lowercase letters indicate significant difference (P < 0.05); different capital letters indicate extremely significant difference (P < 0.01). The same applies to the following tables / figures.

[0114] As can be seen from Table 4, throughout the experimental period, the total weight gain and average daily gain (ADG) of Group III were significantly higher than those of Group I and Group II (P < 0.05); the average daily feed intake of beef cattle in Group III was extremely significantly higher than that of Group I and Group II (P < 0.01); the feed-to-weight ratios of Group III were 7.67%, 6.50% and 5.20% lower than those of Group I and Group III, respectively. Therefore, feeding the Chinese herbal medicine feed additive disclosed in Example 1 of the present invention can increase the average daily gain and total weight gain during the whole period; significantly increase the daily dry matter intake of beef cattle, indicating that the Chinese herbal medicine feed additive disclosed in Example 1 promoted the growth performance of beef cattle.

[0115] (2) Determination of digestibility of dietary nutrients

[0116] Determination of conventional nutrient components in feed and feces: CP was determined by referring to the method of "Determination of Crude Protein in Feed" (GB / T6432—2018); NDF was determined by referring to "Determination of Neutral Detergent Fiber in Feed" (GB / T20806—2006) using the filter bag method (semi-automatic fiber analyzer); ADF was determined by referring to the method of "Determination of Acid Detergent Fiber in Feed" (GB / T20805—2006). The specific determination results are shown in Table 5 below:

[0117] Table 5: Effects of fermented compound traditional Chinese medicine on nutrient digestibility of beef cattle (dry matter basis, %)

[0118]

[0119] As can be seen from Table 5, the digestibility of dry matter in Group III during the experimental period was significantly higher than that in Group I, Group II and Group IV (P < 0.05); the results of digestibility of NDF and ADF showed that Group III was significantly higher than Group I, Group II and Group IV (P < 0.05); indicating that the Chinese herbal medicine feed additive disclosed in Example 1 can improve the digestibility of DM, NDF and ADF, and promote the digestion and absorption of nutrients in the diet by beef cattle.

[0120] (3) Determination of sugar, lipid and protein metabolism indexes

[0121] According to the specific instructions of the Lepu (Beijing) kit, triglyceride TG, total protein TP, albumin ALB, total cholesterol TCHO, creatinine CRE, urea nitrogen UREA, and serum glucose GLU were measured using an automatic biochemical analyzer. The specific measurement results are shown in Table 6.

[0122] Table 6 Effects of Invention Example 1 on blood sugar, lipid and protein metabolism indexes of beef cattle

[0123]

[0124] As can be seen from Table 6, the content of TCHO in Group III was significantly lower than that in Group I and Group II during the entire experimental period (P < 0.05), and Group III decreased by 21.12% and 20.56% compared with Group I and Group II; the content of TP in Group II was significantly higher than that in Group I (P < 0.05), and Group II increased by 3.4% compared with Group III; the content of CRE in Group III and Group IV was significantly lower than that in Group I and Group II (P < 0.05), and there were no significant differences among the other treatments for each index. The experimental results show that feeding the Chinese herbal medicine feed additive disclosed in Example 1 of the present invention can promote protein synthesis metabolism in the body, enhance the deposition of fat and nitrogen, indicating that the Chinese herbal medicine feed additive disclosed in Example 1 of the present invention can improve and stabilize the serum biochemical indexes of beef cattle.

[0125] (4) Determination of liver function indexes

[0126] According to the specific instructions of the Lepu (Beijing) kit, the activities of alkaline phosphatase ALP, aspartate aminotransferase AST, and alanine aminotransferase ALT were measured using an automatic biochemical analyzer, and the specific measurement results are shown in Table 7 below:

[0127] Table 7 Effects of the medicine in Example 1 of the invention on the blood liver function indexes of beef cattle

[0128]

[0129] As can be seen from Table 7, the contents of ALT and AST in Group III were significantly lower than those in Group I (P < 0.05). The content of ALT in Group III decreased by 27.8%, 25.8%, and 23.3% compared with Group I, Group II, and Group IV, respectively. The content of AST in Group III decreased by 16.3%, 15.8%, and 11.2% compared with Group I, Group II, and Group IV, respectively. Aspartate aminotransferase AST and alanine aminotransferase ALT generally exist in liver cells. When liver cells are damaged and destroyed, the two enzymes AST and ALT in liver cells enter the blood, and the increase in AST and ALT in the blood indicates liver cell damage. The decrease in the serum contents of AST and ALT in the group fed with the Chinese herbal medicine feed additive disclosed in Example 1 of the present invention indicates that the Chinese herbal medicine feed additive disclosed in Example 1 of the present invention can reduce liver damage.

[0130] (5) Determination of immunoglobulins and immune cell molecules

[0131] According to the specific instructions of the Wuhan Gene Beauty Box ELISA kit, the experiment was carried out to measure the contents of immunoglobulins IgA, IgM, IgG and immune cell molecules CD4+, CD8+ respectively.

[0132] Table 8 Effects of Example 1 of the invention on the immunoglobulins in the blood of beef cattle

[0133]

[0134]

[0135] As can be seen from Table 8, at the end of the entire experimental period, the contents of immunoglobulin A (IgA) and immunoglobulin G (IgG) in the blood of Group III were significantly higher than those of Group I (P < 0.01), and the content of immunoglobulin M (IgM) in Group III was significantly higher than that of Group I (P < 0.05). Immunoglobulins represent an important part of the humoral immunity of animals and have biological functions such as activating antigens and activating complements. Their content in peripheral blood can reflect the immune function status of the animal body, and an increase in content indicates enhanced humoral immunity. This shows that the Chinese herbal medicine feed additive disclosed in Example 1 of the present invention can enhance the humoral immune function of beef cattle.

[0136] The content of CD4+ in Group III and Group IV was extremely significantly higher than that of Group I (P < 0.05). The concentration of CD8+T in the serum of Group III was significantly lower than that of Group I (P < 0.05). When the concentration of CD4+T cells in the serum is relatively high, it indicates that the immune function of the animal body is strong and it is not easily infected with diseases. The main function of CD8+T cells is to eliminate diseased cells in the body. When the concentration of CD8+T cells in the serum is high, it shows that the function of the animal body's own immune system is decreasing and it is unable to maintain the health of the body. This shows that the Chinese herbal medicine feed additive disclosed in Example 1 of the present invention can enhance the immune ability of the body.

[0137] Feeding Experiment 2:

[0138] 1. Experimental Animals

[0139] Forty 3-month-old lambs with a body weight of 25 ± 2.06 kg were selected as experimental animals. They were uniformly immunized, dewormed, and housed individually in pens. They were fed twice a day at 8:00 and 18:00, with free access to food and water. The pre-feeding period was 15 days, and the experimental period was 60 days.

[0140] 2. Composition of the Basal Diet

[0141] The design of the experimental basal diet referred to the Chinese Feeding Standard for Mutton Sheep (NY / T816 - 2004). The composition of the diet and its nutrient levels are shown in Table 9.

[0142] Table 9 Composition and Nutrient Levels of the Basal Diet (Dry Matter Basis)

[0143] Raw materials Content <![CDATA[Nutrient levels 2) > Content Mixed pasture 30.0 Metabolizable energy ME / (MJ / kg) 10.4 Corn 53.0 Crude protein CP 14.4 Soybean meal 14.9 Calcium Ca 0.7 <![CDATA[Calcium hydrogen phosphate CaHPO4]]> 0.1 Phosphorus P 0.3 Table salt NaCl 0.5 Neutral detergent fiber NDF 23.3 <![CDATA[Premix 1) > 0.5 Acid detergent fiber ADF 15.0 <![CDATA[Sodium bicarbonate NaHCO3]]> 1.0 Non-structural carbohydrate NSC 49.9 Total 100.0

[0144] 1) The premix provided per kilogram of the diet: Ca: 1.52 g, P: 0.41 g, Fe: 25 mg, Zn: 35 mg, Cu: 9 mg, Co: 0.1 mg, I: 0.9 mg, Se: 0.25 mg, Mn: 19.5 mg, niacin: 60 mg, VE: 15 IU, VA: 3000 IU, VD3: 1000 IU.

[0145] 2) The metabolic energy is a calculated value, and the other nutritional levels are measured values.

[0146] 3. Experimental design

[0147] Forty 3-month-old lambs with a body weight of 26.37 ± 2.29 kg were selected as experimental animals and randomly divided into 4 groups with 10 lambs in each group. The CON group was the blank control group fed with the basal diet, the T1 group was fed with the feed supplemented with the Chinese herbal medicine feed additive prepared in Comparative Example 5 at 0.2% of the weight of the basal diet, the T2 group was fed with the feed supplemented with the Chinese herbal medicine feed additive disclosed in Example 1 of the present invention at 0.2% of the weight of the basal diet, and the T3 group was fed with the feed supplemented with the untreated Chinese herbal medicine feed additive at 0.2% of the weight of the basal diet (the preparation method of the untreated Chinese herbal medicine feed additive is: selecting the Chinese medicine raw materials in the same proportion by weight as in Example 1, grinding and pulverizing them, passing through a 40-mesh sieve, and directly adding the obtained Chinese medicine raw material powder to the feed); the experimental period was 60 days.

[0148] 4. Collection of samples

[0149] The fecal samples and feed samples were dried in a constant temperature drying oven at 65 °C until constant weight, allowed to return to room temperature for 24 h, weighed and recorded, pulverized with a small universal pulverizer, and sieved and then sub-packed. Collection and pretreatment of fecal samples: In the middle of the experiment, when the formal experiment reached 30 days, 100 g of fecal samples were collected from each sheep before morning feeding every day for 10 consecutive days. The fecal samples collected every day were divided into two parts. One part was added with 10% H2SO4 for nitrogen fixation for crude protein determination, and the other part was stored frozen at -20 °C. At the end of the experiment, all the fecal samples collected from each sheep in 10 days were mixed. After taking a part and drying it at 65 °C to determine the initial moisture, it was pulverized with a Chinese medicine pulverizer and passed through a 40-mesh sieve to obtain the fecal sample analysis sample for routine nutrient component analysis.

[0150] Preparation of serum: Before morning feeding at the end of the formal test period, 5 ml of blood was collected from the jugular vein of the experimental sheep in each group, taking care to avoid shaking to prevent hemolysis. Then it was centrifuged at a low speed of 4000 r / min for 10 min, and the serum was transferred and sub-packed in 1.5 ml enzyme-free and sterile centrifuge tubes and stored frozen at -80 °C for later measurement.

[0151] 5. Determination indexes

[0152] (1) Determination of the growth performance of lambs

[0153] The experimental lambs were weighed on an empty stomach on the 0th day and the 60th day of the experiment to record the initial and final body weights. The daily feed intake was also monitored. After the experiment, the dry matter intake (DMI), average daily gain (ADG), and feed conversion rate were calculated.

[0154] Average daily feed intake (ADFI) = Total feed intake / Total number of experimental days

[0155] Average daily gain (ADG) = Total weight gain during the experimental period / Number of experimental days

[0156] Feed conversion ratio (F / G) = Daily feed weight / Daily feed intake / Daily gain

[0157] Table 10 Experimental data on the growth performance of lambs

[0158]

[0159] Note: In the same row, if there is no letter or data superscript, or if the same letter is included, it indicates no significant difference (P > 0.05); if different lowercase letters are included, it indicates a significant difference (P < 0.05); if different capital letters are included, it indicates a highly significant difference (P < 0.01). The same applies to the following tables / figures.

[0160] As can be seen from Table 10, throughout the experimental period, the final weight of Group T2 was significantly higher than that of the CON group and Group T1 (P < 0.05); the average daily gain (ADG) of Groups T1, T2, and T3 was significantly higher than that of the CON group (P < 0.05); the feed conversion ratio of Group T2 was significantly lower than that of the CON group (P < 0.05). Therefore, feeding the Chinese herbal medicine feed additive disclosed in Example 1 of the present invention can increase the average daily gain and total weight gain during the whole period of meat sheep, indicating that the Chinese herbal medicine feed additive disclosed in Example 1 promotes the growth performance of meat sheep.

[0161] (2) Determination of dietary nutrient digestibility

[0162] According to GB / T 6435 - 2014(25), the crude moisture in the feed was determined, and then the dry matter content was calculated. According to GB / T20806 - 2022(26) and NY / T 1459 - 2007(27), the NDF and ADF contents in the feed were determined using a fiber analyzer (ANKOM A200i, USA). According to GB / T 6432 - 2018(28), the crude protein (CP) content was measured using an automatic Kjeldahl nitrogen analyzer (K9860, Hanon Technologies, China).

[0163] Calculate the apparent digestibility of nutrients:

[0164] Apparent digestibility of nutrients (%) = 100% - [100% × (a / b) × (c / d)]

[0165] In the formula, a: AIA content in the feed; b: AIA content in the feces; c: Content of the nutrient to be measured in the feces; d: Content of the nutrient to be measured in the feed.

[0166] Table 11 Effects of fermented compound Chinese medicine on the nutrient digestibility of lambs (dry matter basis, %)

[0167]

[0168] Note: When there are no letters or numerical superscripts in the same row and the same letter is included, it indicates that the difference is not significant (P > 0.05); when different lowercase letters are included, it indicates that the difference is significant (P < 0.05).

[0169] As can be seen from Table 11, the digestibility of dry matter in the T2 group during the experimental period was significantly higher than that in the CON and T1 groups (P < 0.05); the results of the digestibility of NDF and ADF showed that the NDF in the T2 group was significantly higher than that in the CON, T1, and T3 groups (P < 0.05); it shows that the Chinese herbal medicine feed additive disclosed in Example 1 can improve the digestibility of DM and NDF, and promote the digestion and absorption of nutrients in the diet by lambs.

[0170] (3) Determination of blood biochemical indexes

[0171] According to the specific instructions of the Lepu (Beijing) kit, triglyceride TG, total protein TP, albumin ALB, total cholesterol TCHO, and serum glucose GLU were measured using an automatic biochemical analyzer.

[0172] Table 12 Effects of Example 1 of the present invention on the blood biochemical indexes of lambs

[0173]

[0174] Note: When there are no letters or numerical superscripts in the same row and the same letter is included, it indicates that the difference is not significant (P > 0.05); when different lowercase letters are included, it indicates that the difference is significant (P < 0.05), the same applies to the following tables / figures.

[0175] As can be seen from Table 12, during the entire experimental period, the content of GLU in the T2 group was significantly higher than that in the CON, T1, and T3 groups (P < 0.05); the content of TG in the T2 group was significantly higher than that in the CON, T1, and T3 groups, and the content of TG in the T1 and T3 groups was significantly higher than that in the CON group (P < 0.05); the content of TP in the T2 group was significantly higher than that in the CON group (P < 0.05), and there was no significant difference in other indexes among the remaining treatments. The experimental results show that feeding the Chinese herbal medicine feed additive disclosed in Example 1 of the present invention can promote the synthesis and metabolism of proteins in the body, enhance the deposition of fat and nitrogen, indicating that the Chinese herbal medicine feed additive disclosed in Example 1 of the present invention can improve the serum biochemical indexes of lambs.

[0176] (4) Determination of liver function indexes

[0177] According to the specific instructions of the Lepu (Beijing) kit, the activities of alkaline phosphatase ALP, aspartate aminotransferase AST, and alanine aminotransferase ALT were measured using an automatic biochemical analyzer.

[0178] Table 13 Effects of the drug pair in Example 1 of the present invention on the blood liver function indexes of lambs

[0179]

[0180] Note: In the same row, if there is no letter or data superscript and the same letter is included, it indicates that the difference is not significant (P>0.05); if different lowercase letters are included, it indicates that the difference is significant (P<0.05).

[0181] As can be seen from Table 13, the contents of ALP and AST in the T2 group were significantly lower than those in the CON and T1 groups (P<0.05), and the content of ALT in the T2 group was significantly lower than that in the CON group (P<0.05). Alkaline phosphatase ALP, aspartate aminotransferase AST, and alanine aminotransferase ALT generally exist in hepatocytes. When hepatocytes are damaged and destroyed, the two enzymes AST and ALT in the hepatocytes enter the blood, and the ALP, AST, and ALT in the blood increase, indicating that the hepatocytes are damaged. The serum contents of ALP, AST, and ALT in the group adding the Chinese herbal medicine feed additive disclosed in Example 1 of the present invention decreased, indicating that the Chinese herbal medicine feed additive disclosed in Example 1 of the present invention can reduce liver damage.

[0182] (5) Determination of immunoglobulins

[0183] The test operations were carried out according to the specific instructions of the Wuhan Gene Beauty Box ELISA kit to measure the contents of immunoglobulins IgA, IgM, and IgG respectively.

[0184] Table 14 Effects of Example 1 of the present invention on the immunoglobulins in the blood of beef cattle

[0185]

[0186] Note: In the same row, if there is no letter or data superscript and the same letter is included, it indicates that the difference is not significant (P>0.05); different capital letters indicate that the difference is extremely significant (P<0.01), the same as the following table / figure.

[0187] As can be seen from Table 14, at the end of the entire test period, the content of immunoglobulin A (IgA) in the blood of the T2 group was significantly higher than that in the CON, T1, and T3 groups (P<0.05), and the contents of immunoglobulin G (IgG) and immunoglobulin M (IgM) in the blood of the T2 group were significantly higher than that in the CON group (P<0.05). Immunoglobulins represent an important part of the humoral immunity of animals and have biological functions such as activating antigens and activating complements. The content in peripheral blood can reflect the immune function status of the animal body, and an increase in content indicates enhanced humoral immunity. It shows that the Chinese herbal medicine feed additive disclosed in Example 1 of the present invention can enhance the humoral immune function of lambs.

Claims

1. A preparation method of a Chinese herbal medicine feed additive for improving the production performance of cattle and sheep, characterized in that, It includes the following steps: S1: Drying and pulverizing raw materials: Prepare hawthorn, astragalus membranaceus, atractylodes macrocephala, codonopsis pilosula, poria cocos, Chinese yam, euryale ferox, tangerine peel, lotus seeds, and licorice as raw materials. Respectively conduct drying treatment on each raw material, and then conduct pulverizing treatment on each dried raw material. After pulverizing into fine powder, sieve through a 40-mesh sieve to obtain the raw material powder of each raw material respectively; S2: Mixing raw materials: Weigh the raw material powder of each raw material in step S1 respectively and mix them to obtain a mixed raw material powder; The raw material powder of each raw material is calculated by weight fraction as follows: 2 - 4 parts of hawthorn, 2 - 4 parts of astragalus membranaceus, 1 - 3 parts of atractylodes macrocephala, 1 - 3 parts of codonopsis pilosula, 1 - 3 parts of poria cocos, 1 - 3 parts of Chinese yam, 1 - 3 parts of euryale ferox, 1 - 2 parts of tangerine peel, 1 - 2 parts of lotus seeds, and 1 - 2 parts of licorice. S3: Ultrasonic treatment: Mix the raw material powder after mixing in step S2 with distilled water, transfer the mixed slurry to an ultrasonic cell disruptor for ultrasonic treatment to obtain an ultrasonically treated slurry; S4: Enzymatic hydrolysis treatment: Add a complex enzyme to the ultrasonically treated slurry obtained in step S3 for enzymatic hydrolysis to obtain an enzymatically hydrolyzed mixture; S5: Fermentation treatment: Dehydrate and dry the enzymatically hydrolyzed mixture obtained in step S4 to obtain an enzymatically hydrolyzed dry powder. Place the enzymatically hydrolyzed dry powder into a fermentation container, add water and mix evenly, then add a complex bacterial solution for fermentation. After fermentation is completed, a Chinese herbal medicine feed additive is obtained.

2. The preparation method of a Chinese herbal medicine feed additive for improving the production performance of cattle and sheep according to claim 1, characterized in that, The drying conditions in step S1 are drying at 60°C - 65°C for 4 - 8 hours.

3. The preparation method of a Chinese herbal medicine feed additive for improving the production performance of cattle and sheep according to claim 1, characterized in that, In step S3, the raw material powder and distilled water are mixed at a liquid - solid ratio of 10∶1.

4. The preparation method of a Chinese herbal medicine feed additive for improving the production performance of cattle and sheep according to claim 1, characterized in that, The conditions for ultrasonic treatment of the ultrasonic cell disruptor in step S3 are an ultrasonic power of 400 - 600W and an ultrasonic time of 25 - 45min.

5. The preparation method of a Chinese herbal medicine feed additive for improving the production performance of cattle and sheep according to claim 1, characterized in that, The complex enzyme in step S4 is a complex enzyme preparation composed of pectinase > 30U / g, xylanase > 1200U / g, β - glucanase > 200U / g, and cellulase > 30U / g.

6. The preparation method of a Chinese herbal medicine feed additive for improving the production performance of cattle and sheep according to claim 5, characterized in that, The added mass of the complex enzyme in step S4 is 0.2 - 0.5% of the mass of the mixed raw material powder in step S2, the temperature for enzymatic hydrolysis is 30 - 50°C, and the enzymatic hydrolysis treatment time is 4 - 6h.

7. The preparation method of a Chinese herbal medicine feed additive for improving the production performance of cattle and sheep according to claim 1, characterized in that, The mass of water added in step S5 is 20 - 50% of the mass of the enzymatically hydrolyzed dry powder.

8. The preparation method of a Chinese herbal medicine feed additive for improving the production performance of cattle and sheep according to any one of claims 7, characterized in that, The complex bacterial solution in step S5 is a complex bacterial solution of bacillus subtilis and bacillus licheniformis.

9. The preparation method of a Chinese herbal medicine feed additive for improving the production performance of cattle and sheep according to claim 8, characterized in that, The fermentation conditions in step S5 are: the added mass of the complex bacterial solution is 3 - 5% of the mass of the enzymatically hydrolyzed dry powder, and the fermentation days are 3 - 5 days.

10. A Chinese herbal medicine feed additive for improving the production performance of cattle and sheep, characterized in that, It is prepared by the preparation method of the Chinese herbal medicine feed additive for improving the production performance of cattle and sheep according to any one of claims 1 - 9.

Images