A feed additive prepared based on traditional Chinese medicine residues, a preparation method and application thereof

Abstract

The application provides a feed additive prepared based on traditional Chinese medicine residues, and a preparation method and application thereof. The feed additive is prepared from a mixture of traditional Chinese medicine residues, vinasse, glutamic acid residues, bran and puffed corn flour after fermentation by spherical lysinibacillus; the spherical lysinibacillus (Lysinibacillus sphaericus) LD-LS-156 is preserved in the China Center for Type Culture Collection of Wuhan University, and the preservation number is CCTCC M 2023775. The feed additive added into feed can effectively improve the egg production rate, improve the egg quality, and reduce the diarrhea rate. The application utilizes industrial waste such as traditional Chinese medicine residues and vinasse to prepare the feed additive through fermentation, changes waste into treasure, solves pollution, widens the development and reuse of unconventional feed resources, and provides a new idea for relieving the increasingly insufficient conventional feed resources.

Description

Technical Field

[0001] This invention belongs to the field of feed additive technology, specifically relating to a feed additive prepared from traditional Chinese medicine residue, its preparation method, and its application. Background Technology

[0002] With the continuous expansion of my country's livestock industry, conventional feed resources are becoming increasingly scarce. Coupled with rising consumer demands for food safety and the growing contradiction between the overuse of antibiotics and the resulting harm to animal health, the feed industry faces urgent challenges related to food safety, grain shortages, and environmental pollution. Bio-feed, an emerging industry, transforms feed ingredients into microbial cell protein, bioactive small peptides and amino acids, active probiotics, and compound enzyme preparations—materials that are more easily consumed, digested, and absorbed by livestock and poultry, resulting in higher nutritional value and no toxicity. Therefore, bio-fermentation technology can increase the added value of unconventional raw materials, broaden the scope of development and reuse of unconventional feed resources, reduce breeding costs, ensure the quality and safety of animal products, and reduce environmental pollution from livestock farming, thereby promoting the healthy and sustainable development of the feed industry. Summary of the Invention

[0003] In view of this, the present invention provides a feed additive based on traditional Chinese medicine residue, its preparation method and application.

[0004] To achieve the above objectives, the present invention adopts the following technical solution:

[0005] A feed additive based on traditional Chinese medicine residue, wherein the feed additive is prepared by fermentation of a mixture of traditional Chinese medicine residue, distiller's grains, glutamic acid residue, wheat bran, and puffed corn flour by Bacillus spheroidae.

[0006] The spherical lysinibacillus sphaericus LD-LS-156 is deposited at the China Center for Type Culture Collection, Wuhan University, with accession number CCTCC M2023775.

[0007] Furthermore, the fermenting bacteria in the fermentation process also include at least one of Bacillus subtilis and Saccharomyces cerevisiae.

[0008] Furthermore, the moisture content of the medicinal herb residue is 40%–60%, and the moisture content of the distiller's grains is 40%–60%.

[0009] In some specific embodiments, preferably, the moisture content of the Chinese herbal medicine residue is 50%, and the moisture content of the distiller's grains is 50%.

[0010] Furthermore, the mass ratio of Chinese herbal medicine residue, distiller's grains, and wheat bran in the mixture is (4-6):(2.5-3.5):(1.5-2.5).

[0011] In some specific embodiments, preferably, the mass ratio of Chinese herbal medicine residue, distiller's grains, and wheat bran in the mixture is 5:3:2.

[0012] Furthermore, the amount of glutamic acid residue and puffed corn flour added to the mixture is 0.5% to 1.5% of the sum of the mass of Chinese herbal medicine residue, distiller's grains, and wheat bran.

[0013] In some specific embodiments, preferably, the amount of glutamic acid residue and puffed corn flour added to the mixture is 1% of the sum of the mass of Chinese herbal medicine residue, distiller's grains, and wheat bran.

[0014] Furthermore, the medicinal residue includes astragalus residue, angelica residue, codonopsis residue, wolfberry residue, jujube residue, licorice residue, cinnamon residue, clove residue, epimedium residue, and mulberry leaf residue.

[0015] Furthermore, the mass ratio of Astragalus membranaceus residue, Angelica sinensis residue, Codonopsis pilosula residue, Lycium barbarum residue, Jujube residue, Glycyrrhiza uralensis residue, Cinnamomum cassia residue, Clove residue, Epimedium brevicornu residue, and Mulberry leaf residue in the Chinese herbal medicine residue is 0.8:1:0.5:1:1:0.2:0.2:0.1:0.1:0.5.

[0016] A method for preparing the above-mentioned feed additive includes the following steps:

[0017] Prepare a bacterial seed solution containing Bacillus spheroidans;

[0018] Mix Chinese medicinal herb residue, distiller's grains, glutamic acid residue, wheat bran, and puffed corn flour evenly, inoculate with the above-mentioned live bacteria seed liquid, and dry, crush, and sieve the fermented material to obtain the feed additive.

[0019] Furthermore, the seed liquid also includes a brewer's yeast seed liquid;

[0020] Alternatively, the fermentation process may also involve inoculation with Bacillus subtilis powder;

[0021] Alternatively, the seed liquid may also include a brewer's yeast seed liquid, and the fermentation may also be inoculated with Bacillus subtilis powder.

[0022] Furthermore, the inoculation mass of the seed liquid accounts for 2% to 4% of the mass of the mixture to be fermented, and the inoculation mass of the Bacillus subtilis powder accounts for 0.08% to 0.1% of the mass of the mixture to be fermented.

[0023] When the inoculated seed liquid is Bacillus spheroides seed liquid or Saccharomyces cerevisiae seed liquid, the inoculation mass ratio of Bacillus spheroides seed liquid to Saccharomyces cerevisiae seed liquid is 1:1.

[0024] Fermentation conditions: temperature 28℃, time 50h, ventilation and temperature control during fermentation, material temperature not exceeding 35℃.

[0025] A chicken feed, wherein the above-mentioned feed additives are added at a mass fraction of 1% to 2%.

[0026] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0027] (1) This invention utilizes industrial waste such as Chinese medicine residue and distiller's grains to prepare feed additives through fermentation, turning waste into treasure, solving pollution problems, and expanding the development and reuse of unconventional feed resources, providing a new approach to alleviate the increasing scarcity of conventional feed resources.

[0028] (2) The feed additive prepared by this invention makes full use of the active ingredients such as polysaccharides, saponins, alkaloids and flavonoids in Chinese herbal medicine residues, as well as the rich amino acids, vitamins and proteins in distiller's grains; it can effectively alleviate the diarrhea rate of laying hens, effectively reduce harmful bacteria such as Escherichia coli and Salmonella, and at the same time increase the content of beneficial bacteria such as Lactobacillus and Bifidobacterium; in addition, it can significantly improve the quality of eggs, improve the yolk color, increase the eggshell thickness, the laying rate and the average egg weight.

[0029] (3) In this invention, Bacillus spheroidae can produce a variety of hydrolytic enzymes, which can effectively degrade crude fiber, protein and other macromolecules in Chinese medicine residues. At the same time, it also has antibacterial effects and can effectively inhibit diarrhea, infection and other problems caused by pathogenic bacteria such as Escherichia coli and Salmonella in the animal intestines. When Bacillus subtilis and Saccharomyces cerevisiae are combined and applied to Chinese medicine residues, they can degrade into small molecule active substances, enrich the diet of the fed animals, improve palatability, and at the same time, the microorganisms colonize in the intestines, inhibit harmful bacteria and promote beneficial bacteria, regulate the intestinal flora structure, improve intestinal health, stabilize the intestinal environment, and thus promote the digestion and absorption of nutrients by the body. When fermentation is carried out using mixed strains, the effect is even better. Detailed Implementation

[0030] The present invention will now be described in further detail with reference to specific embodiments, so that those skilled in the art can more clearly understand the present invention.

[0031] Key experimental material sources and physicochemical parameters:

[0032] The spherical lysine-containing Bacillus was provided by Lude Environmental Technology Co., Ltd., with accession number: CCTCC M2023775;

[0033] Bacillus subtilis (1×10) 11 cfu / g), Saccharomyces cerevisiae (2×10) 10 All CFU / g were purchased from Shandong Weilan Biotechnology Co., Ltd.

[0034] The Chinese herbal medicine residue is the residue produced by a biomedical company in Ezhou, Hubei Province, after digital extraction. It mainly includes residues of Astragalus membranaceus, Angelica sinensis, Codonopsis pilosula, Lycium barbarum, Jujube, Glycyrrhiza uralensis, Cinnamomum cassia, Clove, Epimedium brevicornu, and Mulberry leaf.

[0035] The distiller's grains were supplied by a winery in Bozhou, Anhui Province.

[0036] The glutamic acid residue was supplied by a food ingredient manufacturer in Heze, Hebei Province.

[0037] All other raw materials or structures not specifically described in this invention already exist in the prior art and can be purchased directly from the market.

[0038] Example 1

[0039] This embodiment provides a feed additive, and the specific preparation steps are as follows:

[0040] S1. Take the astragalus residue, angelica residue, codonopsis residue, wolfberry residue, jujube residue, licorice residue, cinnamon residue, clove residue, epimedium residue, and mulberry leaf residue and mix them evenly in a mass ratio of 0.8:1:0.51:1:0.2:0.2:0.1:0.1:0.5 to obtain the medicinal residue premix.

[0041] Mix the medicinal residue premix (50% moisture content), distiller's grains (50% moisture content), and wheat bran in a mass ratio of 5:3:2. Then add glutamic acid residue and puffed corn flour at 1% of the mass of the mixture, mix evenly, and you will get the mixture to be fermented.

[0042] S2. Inoculate *Bacillus spheroides* slant culture onto seed culture medium (10 g / L peptone, 5 g / L yeast extract, 10 g / L sodium chloride, pH adjusted to 7.0 with 5 mol / L NaOH, and brought to a final volume of 1 L with deionized water; autoclave at 121℃ for 20 min). Activate the medium in a shaker at 37℃ for 18 h to prepare primary seed culture. Then, inoculate at a 2% inoculum and culture for 24 h to obtain activated *Bacillus spheroides* seed culture, with viable cell count controlled at 4 × 10⁻⁶. 8 cfu / mL;

[0043] Inoculate 2% of the yeast powder into a 2% sucrose solution and culture at 28°C for 1 hour to obtain activated yeast seed culture.

[0044] S3. Inoculate the activated Bacillus spheroides seed liquid, Saccharomyces cerevisiae seed liquid, and Bacillus subtilis powder into the mixture to be fermented. The inoculation amount is calculated based on 100g of the mixture to be fermented. The inoculation amount of Bacillus spheroides seed liquid and Saccharomyces cerevisiae seed liquid is 1.5g each, and the inoculation amount of Bacillus subtilis powder is 0.09g. After mixing evenly, place it at 28℃ for 50h for fermentation. The temperature is controlled by blowing air during the fermentation process, and the material temperature does not exceed 35℃ to obtain the fermented material.

[0045] S4. Dry the above fermented material at 65℃ until the moisture content is about 30%. After drying, crush it with a pulverizer and pass it through a 10-mesh sieve to obtain the feed additive.

[0046] Example 2

[0047] This embodiment provides a feed additive, whose raw materials and steps are basically the same as those in Embodiment 1, except that: in step S1, the medicinal residue premix, distiller's grains, and wheat bran are added in a mass ratio of 4:3.5:1.5, and the amount of glutamic acid residue and puffed corn flour added is 0.5% of the mass of the mixture; in step S3, the inoculation amount is calculated based on 100g of the mixture to be fermented, and the inoculation amount of Bacillus spheroides seed liquid and Saccharomyces cerevisiae seed liquid is 1g, and the inoculation amount of Bacillus subtilis powder is 0.1g.

[0048] Example 3

[0049] This embodiment provides a feed additive, whose raw materials and steps are basically the same as those in Embodiment 1, except that: in step S1, the medicinal residue premix, distiller's grains, and wheat bran are added in a mass ratio of 6:2.5:2.5, and the amount of glutamic acid residue and puffed corn flour added is 1.5% of the mass of the mixture; in step S3, the inoculation amount is calculated based on 100g of the mixture to be fermented, and the inoculation amount of Bacillus spheroides seed liquid and Saccharomyces cerevisiae seed liquid is 2g, and the inoculation amount of Bacillus subtilis powder is 0.08g.

[0050] Example 4

[0051] This embodiment provides a feed additive whose raw materials and steps are basically the same as those in Example 1, except that: in step S3, only activated Bacillus spheroidans seed liquid is used for fermentation, and at the same time, an equal amount of 2% sucrose solution as in the Saccharomyces cerevisiae seed liquid in Example 1 is added.

[0052] Example 5

[0053] This embodiment provides a feed additive whose raw materials and steps are basically the same as those in Example 1, except that: in step S3, a combination of Bacillus spheroidans and Bacillus subtilis seed liquid is used for fermentation, and at the same time, an equal amount of 2% sucrose solution is added to the seed liquid of Saccharomyces cerevisiae in Example 1.

[0054] Example 6

[0055] This embodiment provides a feed additive, whose raw materials and steps are basically the same as those in Embodiment 1, except that: in step S3, a combination of Bacillus spheroidans and Saccharomyces cerevisiae seed liquid is used for fermentation.

[0056] Comparative Example 1

[0057] This comparative example provides a feed additive whose raw materials and steps are basically the same as those in Example 1, except that: in step S3, a combination of Bacillus subtilis and Saccharomyces cerevisiae seed liquid is used for fermentation, the seed liquid of Bacillus spheroides lysine is removed, and an amount of sterilized seed liquid culture medium for activation is added at the same time as the Bacillus spheroides lysine seed liquid in Example 1.

[0058] Comparative Example 2

[0059] This comparative example provides a feed additive whose raw materials and steps are basically the same as those in Example 1, except that: in step S3, the brewer's yeast seed liquid is replaced by Aspergillus niger powder, and at the same time, an amount of sterilized seed liquid culture medium for activation is added, which is the same as that of Bacillus spheroidans seed liquid in Example 1.

[0060] To further determine the changes in nutritional indicators, tests were conducted before and after fermentation for Examples 1 and 4-6, and the results are shown in Table 1.

[0061] Table 1 Nutritional indicators before and after fermentation in the examples

[0062]

[0063] Table 1 shows that the content of nutrients such as true protein and crude fat increased to some extent after fermentation, indicating that microbial fermentation helps to improve the nutritional components of the material. At the same time, the crude fiber content decreased to varying degrees, improving palatability and facilitating absorption. Comparative analysis shows that when three strains of Bacillus spheroidans seed liquid, Bacillus subtilis, and Saccharomyces cerevisiae were fermented together, the proportion of true protein increased the highest, the crude fiber content was lower, and the overall fermentation effect was the best.

[0064] Further research was conducted on the effects of adding the feed additives prepared in Examples 1 and 4, and Comparative Examples 1-2 to the feed on laying hens. The following experiments were also carried out at a laying hen farm in Xiaogan City, Hubei Province.

[0065] Seventy-two hundred Hy-Line Brown chickens aged 50 weeks in good health were selected and randomly divided into groups. Each group had six replicates, with 20 animals per replicate, and four chickens per cage. The feeding period was 70 days, with a 10-day acclimatization period, and the experimental period was 60 days. During the acclimatization period, all groups were fed normal complete chicken feed, which was gradually replaced with commercial feed over 10 days. Specific grouping and feeding arrangements are shown in Table 2 below.

[0066] Table 2 Specific groupings and feeding arrangements

[0067]

[0068]

[0069] During the experiment, the chicken coop was kept well-ventilated, with supplemental lighting in the morning and evening to ensure adequate illumination. The room temperature was maintained at 22-25℃, and the relative humidity was 60-75%. Feeding was conducted three times a day at fixed times, and immunization and deworming were carried out as usual. Dedicated personnel managed the chickens, observing and recording the growth performance and egg production of each group. Eggs were collected twice daily to minimize stress.

[0070] After feeding, the diarrhea rate and fecal microorganisms of each group of laying hens were statistically analyzed as shown in Table 3 below:

[0071] Table 3. Diarrhea rate and fecal microbial statistics of laying hens in each group.

[0072]

[0073] The Kjeldahl method was used to determine the true protein content.

[0074] As shown in Table 3, the addition of feed additives prepared in different embodiments or comparative proportions can reduce the diarrhea rate of laying hens to varying degrees. When the addition amount in the embodiment is 2%, the diarrhea rate can be reduced, and harmful bacteria such as Escherichia coli and Salmonella can also be appropriately reduced. At the same time, the content of beneficial bacteria such as Lactobacillus and Bifidobacterium can be increased. The effect is better when the addition amount is 1%.

[0075] The diarrhea rate and microbial indicators were reduced after adding the feed additive prepared in Example 4. Although the effect was not as significant as that of compound bacterial fermentation, it still had a positive effect on the reuse of industrial waste such as drug residues and distiller's grains.

[0076] After adding the feed additive prepared in Comparative Example 1, compared with the control group and Example 4, the diarrhea rate of laying hens was 12.99%, which was 1.62% lower than the control group and 1% lower than Example 4. The proportion of Escherichia coli was lower than both, and the proportion of probiotics was appropriately higher than the control group and Example 4. It can be seen that the feeding effect of mixed fermented feed of Bacillus lysine and Bacillus subtilis is better than that of single fermented feed of Bacillus spheroidae.

[0077] After adding the feed additive prepared in Comparative Example 2, the diarrhea rate was reduced to some extent compared with the control group, but the intestinal pathogens were not improved. Compared with the control group, the number of Escherichia coli was almost the same, while Salmonella increased to some extent, with an increase of 1.52 cfu / g. Probiotics also increased to some extent, but the proportion was very small, indicating that the feeding effect of Aspergillus niger fermented feed was not good.

[0078] Further statistical analysis was conducted on the growth and egg production of each group of laying hens, and the results are shown in Table 4:

[0079] Table 4. Statistics on the growth and egg production of laying hens in each group.

[0080]

[0081] Among them, the EMT-7300 multi-functional egg product analyzer detects the color of egg yolks.

[0082] As shown in Table 4, the addition of the feed additive prepared in Example 1 can significantly improve the growth performance of laying hens, improve the yolk color of the eggs, and increase the eggshell thickness, laying rate, and average egg weight. Based on production performance, the effect is relatively better when the addition amount is 1%, while compared with egg quality, the effect is better when the addition amount is 2%.

[0083] After adding the feed additive prepared in Example 4, the laying hen production performance and egg quality were slightly improved compared with the control group; although the effect was not as significant as that of compound bacterial fermentation, it still had a positive effect on the reuse of industrial waste such as drug residues and distiller's grains.

[0084] After adding the feed additive prepared in Comparative Example 1, compared with the control group, the egg production rate, feed conversion ratio and average egg weight all decreased slightly, but the feed intake increased to a certain extent, and the yolk color improved to a certain extent.

[0085] After adding the feed additive prepared in Comparative Example 2, the egg production rate and feed conversion ratio both decreased slightly compared with the control group, but the average egg weight and feed intake increased to some extent, and the yolk color improved to some extent.

[0086] Considering factors such as the nutritional composition of fermented feed, diarrhea rate, fecal microorganisms, production performance, and egg quality, the best feeding effect is achieved by compound fermented feed containing Bacillus spheroides lysine seed liquid, Bacillus subtilis, and Saccharomyces cerevisiae.

[0087] The above are merely preferred embodiments of the present invention and are not intended to limit the scope of protection of the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A chicken feed additive prepared based on traditional Chinese medicine residue, characterized in that, The chicken feed additive is prepared by fermentation of a mixture of Chinese herbal medicine residue, distiller's grains, glutamic acid residue, wheat bran, and puffed corn flour by Bacillus spheroidae. The spherical lysine-containing Bacillus is Lysinibacillus sphaericus LD-LS-156, It is deposited at the China Center for Type Culture Collection, Wuhan University, with accession number CCTCC M 2023775; Bacillus subtilis and Saccharomyces cerevisiae were also added to the fermentation process. The medicinal residues mentioned are astragalus residue, angelica residue, codonopsis residue, wolfberry residue, jujube residue, licorice residue, cinnamon residue, clove residue, epimedium residue, and mulberry leaf residue; The mass ratio of medicinal herb residue, distiller's grains, and wheat bran in the mixture is (4~6):(2.5~3.5):(1.5~2.5). The amount of glutamic acid residue and puffed corn flour added to the mixture is 0.5% to 1.5% of the sum of the mass of Chinese herbal medicine residue, distiller's grains, and wheat bran, respectively. The chicken feed additive is added to chicken feed at a rate of 1% to 2%; The moisture content of the medicinal herb residue is 40%~60%, and the moisture content of the distiller's grains is 40%~60%. The mass ratio of Astragalus membranaceus residue, Angelica sinensis residue, Codonopsis pilosula residue, Lycium barbarum residue, Jujube residue, Glycyrrhiza uralensis residue, Cinnamomum cassia residue, Clove residue, Epimedium brevicornu residue, and Mulberry leaf residue in the Chinese herbal medicine residue is 8:10:5:10:10:2:2:1:1:

5.

2. A method for preparing the chicken feed additive as described in claim 1, characterized in that, Includes the following steps: Prepared spherical lysine-containing Bacillus seed culture and Saccharomyces cerevisiae seed culture; Mix Chinese medicinal herb residue, distiller's grains, glutamic acid residue, wheat bran, and puffed corn flour evenly, then inoculate with Bacillus spheroides seed liquid, Saccharomyces cerevisiae seed liquid, and Bacillus subtilis powder for fermentation. Dry, crush, and sieve the fermented material to obtain chicken feed additive.

3. The preparation method according to claim 2, characterized in that, The total inoculation mass of the *Bacillus spheroidans* seed liquid and *Saccharomyces cerevisiae* seed liquid accounts for 2% to 4% of the mass of the mixture to be fermented, and the inoculation mass of the *Bacillus subtilis* powder accounts for 0.08% to 0.1% of the mass of the mixture to be fermented. The inoculation mass ratio of Bacillus spheroidans seed liquid and Saccharomyces cerevisiae seed liquid was 1:

1. Fermentation conditions: temperature 28℃, time 50h, ventilation and temperature control during fermentation, material temperature not exceeding 35℃.

4. A type of chicken feed, characterized in that, The chicken feed contains the chicken feed additive described in claim 1 at a mass fraction of 1% to 2%.

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