Green feed additive and preparation method thereof

Abstract

The application discloses a kind of green feed additive and preparation method thereof, the green feed additive, including the following mass parts of component: sugar terpenoid 30-50 parts, peach leaf extract 8-12 parts and birch truffle extract 0.1-0.5 parts.The application is compounded with sugar terpenoid, peach leaf extract and birch truffle extract, wherein, sugar terpenoid is the natural active substance extracted from camellia plant oil seed cake, peach leaf extract and birch truffle extract are both traditional Chinese medicine extract, the three are compounded, which will not cause bacterial drug resistance, and can reduce the hidden trouble of food safety;Make the weight of broiler chicken increase fast, have obvious weight gain effect to broiler chicken, and feed to weight ratio is reduced by 30.19%, significantly improve the growth performance of broiler chicken;Can enhance immunity, promote antibody secretion, accelerate immune cell proliferation, enhance antigen antibody level, to realize the rapid protection to broiler chicken.

Description

Technical Field

[0001] This invention relates to the technical field of animal feed processing, and specifically to a green feed additive and its preparation method. Background Technology

[0002] The discovery of antibiotics and the successful synthesis of chemical antibacterial substances have played a vital role in the prevention and treatment of animal diseases, improving animal production performance, and increasing livestock product output. However, the widespread use of antibiotics as growth promoters and disease control feed additives has led to a growing awareness of their drawbacks. Long-term antibiotic use has resulted in antibiotic resistance in bacteria. In livestock farming, bacterial diseases such as E. coli, Staphylococcus, and Salmonella, which were previously not serious or rare, have now become major infectious diseases in poultry, directly related to the long-term overuse of antibiotics. Long-term antibiotic use also reduces animal disease resistance and endangers food safety. Antibiotic residues in livestock products directly harm human health. Therefore, in livestock production, there is an increasing preference for using pollution-free and residue-free natural green feed additives.

[0003] Glycoterpenoids are natural active substances extracted from the oilseed cake of Camellia plants. They are my country's first independently researched and developed innovative feed additive product approved by the Ministry of Agriculture and Rural Affairs. Composed of sugars, triterpenoid saponins, and organic acids, they possess anti-stress, immune-enhancing, and animal production performance-improving effects. The promotion and application of glycoterpenoids will undoubtedly play a significant role in advancing the green development of my country's feed industry, addressing drug residue issues, and realizing the greening of animal-derived foods.

[0004] With the emergence of problems such as residues, drug resistance, and toxic side effects on animals from synthetic drugs, antibiotics, and other pharmaceutical additives, both domestically and internationally, the search has begun for safer and more effective feed additives from traditional Chinese medicine. Traditional Chinese medicine is made from natural substances, making it safe and reliable with fewer side effects. Some traditional Chinese medicines themselves are rich in protein, vitamins, and minerals, possessing both medicinal and nutritional functions. Peach leaves, the leaves of the peach or wild peach plant (Rosaceae family), are reported to contain glycosides, naringenin, quinic acid, hydroxyl groups, and various organic acids. They have the effects of dispelling wind and clearing heat, drying dampness and detoxifying, and killing parasites. They can be used for external wind-evil, headache, wind-dampness, eczema, carbuncles, sores, tinea, malaria, and trichomoniasis. Birch polypore (Inonotus obliquus), also known as birch polypore fungus, is a rather rare polypore fungus discovered in northern latitudes since the 16th century. In Nordic countries and Russia, it is considered to have therapeutic effects on tuberculosis, heart disease, and cancer. Birch polypore has a complex composition, containing polysaccharides, triterpenoids, folic acid derivatives, aromatic vanillic acid, and alkaloids, among other active substances, and possesses antioxidant, hypoglycemic, and immunomodulatory effects. Currently, no methods have been reported for the effective utilization of glycoterpenoids, peach leaves, and birch polypore. Summary of the Invention

[0005] In view of the above-mentioned prior art, the purpose of this invention is to provide a green feed additive and its preparation method. The green feed additive of this invention can enhance the growth performance and improve the immunity of broilers.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] In a first aspect, the present invention provides a green feed additive comprising the following components in parts by weight: 30-50 parts of glycoterpenoids, 8-12 parts of peach leaf extract and 0.1-0.5 parts of birch polypore extract.

[0008] Preferably, the green feed additive comprises the following components in parts by weight: 40 parts of glycoterpenoids, 10 parts of peach leaf extract, and 0.2 parts of birch polypore extract.

[0009] Preferably, the peach leaf extract is obtained by the following method:

[0010] Add 80-95% ethanol (by volume) to peach leaf powder, extract 2-4 times with ultrasound, filter, and obtain the extract; concentrate and dry the extract to obtain peach leaf extract.

[0011] Preferably, the ratio of peach leaf powder to ethanol is 1g:(20-30)ml; the ultrasonic extraction time is 10-15min; and the ultrasonic power is 150W-200W.

[0012] Preferably, the birch polypore extract is obtained by the following method:

[0013] (1) Crush the birch polypores and sieve them, then add water as a fermentation medium;

[0014] (2) Inoculate the fermentation medium with a mixed seed liquid, wherein the inoculation amount of the mixed seed liquid is 5-10% of the volume of the fermentation medium, and the mixed seed liquid is composed of Bacillus retroflexus, Candida utilis and Bacillus coagulans; ferment at a constant temperature of 30-35℃ for 60-72h, filter, and obtain fermentation broth; concentrate and dry the fermentation broth to obtain birch polypore extract.

[0015] Preferably, the *Brevibacillus laterosporus* species in question has the accession number CGMCC.

[0016] 1.4826 was purchased from the China General Microbiological Culture Collection Center (CGMCC); the accession number of the *Candida utilis* was CGMCC 2.2876, also purchased from the CGMCC; the accession number of the *Bacillus coagulans* was CGMCC.

[0017] 1.6563, purchased from the China General Microbiological Culture Collection Center of the China Association for the Preservation and Management of Microbial Cultures.

[0018] Preferably, in step (1), the mass ratio of birch polypore to water is 1:15 to 20.

[0019] Preferably, in step (2), the concentration of *Bacillus laterosporus* in the mixed seed solution is 1.0 × 10⁻⁶. 8 ~1.0×10 9 The concentration of *C. utilis* was 1.0 × 10⁻⁶ CFU / mL. 7 ~1.0×10 8 The concentration of Bacillus coagulans was 1.0 × 10⁻⁶ CFU / mL. 7 ~1.0×10 8 cfu / mL.

[0020] A second aspect of the present invention provides a method for preparing the above-mentioned green feed additive, comprising the following steps:

[0021] The glycosides, peach leaf extract, and birch polypore extract were weighed according to their respective weight proportions, mixed evenly, and a green feed additive was obtained.

[0022] The beneficial effects of this invention are:

[0023] 1. This invention combines glycosides, peach leaf extract, and birch polypore extract. Glycosides are natural active substances extracted from the oilseed cake of Camellia plants. Peach leaf extract and birch polypore extract are both traditional Chinese medicine extracts. Combining these three ingredients will not lead to antibiotic resistance in bacteria and can reduce potential food safety risks, providing a green and environmentally friendly natural drug source.

[0024] 2. This invention combines glycosides, peach leaf extract, and birch polypore extract to accelerate the weight gain of broilers, significantly improve their weight gain, and reduce the feed conversion ratio by 30.19%, thus significantly improving their growth performance. It can also enhance immunity, promote antibody secretion, accelerate immune cell proliferation, and enhance antigen-antibody levels to achieve rapid protection for broilers. Detailed Implementation

[0025] It should be noted that the following detailed descriptions are illustrative and intended to provide further explanation of this application. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains.

[0026] As described in the background section, the overuse of antibiotics can lead to antibiotic resistance in bacteria, reducing animal disease resistance and endangering food safety. In livestock and poultry production, there is an increasing preference for using pollution-free and residue-free natural green feed additives. Based on this, the inventors used glycopterpenoids, a green feed additive, as the main ingredient, which enhances immunity and improves animal production performance. Furthermore, the inventors consulted numerous literatures and screened various traditional Chinese medicines with immunomodulatory capabilities, including Astragalus membranaceus, ginseng, Codonopsis pilosula, birch polypore, Atractylodes macrocephala, and Panax quinquefolius. Considering the antibiotic resistance problem, the inventors hypothesized that peach leaves, containing glycosides, naringenin, quinic acid, hydroxyl groups, and various organic acids, possess antibacterial, heat-clearing, and detoxifying effects. The inventors hypothesized that combining peach leaves with the aforementioned immune-enhancing herbal ingredients and glycopterpenoids might yield unexpected results. The inventors combined the above-mentioned Chinese herbal medicine extracts, peach leaf extract, and glycoterpenoids to investigate their effects on the growth performance and immunity of poultry. They found that the combination of birch polypore extract and peach leaf extract could significantly improve the growth performance and immunity of chickens.

[0027] To enable those skilled in the art to better understand the technical solution of this application, the technical solution of this application will be described in detail below with reference to specific embodiments.

[0028] The test materials used in the embodiments of this invention are all conventional test materials in the art and can be purchased through commercial channels.

[0029] The glycoterpenoids used in this invention were purchased from Xiamen Zhongnongke Chemical New Materials Co., Ltd.

[0030] The *Brevibacillus laterosporus* used in this invention has the accession number CGMCC1.4826 and was purchased from the China General Microbiological Culture Collection Center; the *Candidautilis* has the accession number CGMCC2.2876 and was purchased from the China General Microbiological Culture Collection Center; and the *Bacillus coagulans* has the accession number CGMCC1.6563 and was purchased from the China General Microbiological Culture Collection Center.

[0031] Example 1: Preparation of green feed additives:

[0032] 1. Raw material composition (parts by weight): 30 parts of glycoterpenoids, 8 parts of peach leaf extract, and 0.1 parts of birch polypore extract;

[0033] 2. Preparation steps:

[0034] (1) Preparation of peach leaf extract:

[0035] Add 95% ethanol (by volume) to peach leaf powder at a ratio of 1g:25ml; perform ultrasonic extraction four times for 10 minutes each time, using an ultrasonic power of 200W; filter to obtain the extract; concentrate and dry the extract to obtain peach leaf extract.

[0036] (2) Preparation of Chaga mushroom extract:

[0037] After crushing the birch polyp and sieving it, water was added as a fermentation medium, with a mass ratio of birch polyp to water of 1:15.

[0038] A mixed seed culture was inoculated into the fermentation medium at an inoculation volume of 8% of the fermentation medium volume. The mixed seed culture consisted of *Bacillus laterosporus*, *Candida utilis*, and *Bacillus coagulans*. The concentration of *Bacillus laterosporus* in the mixed seed culture was 1.0 × 10⁻⁶. 9 The concentration of *C. utilis* was 1.0 × 10⁻⁶ CFU / mL. 7 The concentration of Bacillus coagulans was 1.0 × 10⁻⁶ CFU / mL. 8 cfu / mL;

[0039] Fermentation was carried out at a constant temperature of 35℃ for 68 hours, and the mixture was filtered to obtain the fermentation broth. The fermentation broth was then concentrated and dried to obtain the birch polypore extract.

[0040] (3) Weigh out the glycosides, peach leaf extract and birch polypore extract according to the mass fraction, mix them evenly, and obtain the green feed additive.

[0041] Example 2: Preparation of green feed additives:

[0042] Raw material composition (parts by weight): 40 parts of glycoterpenoids, 10 parts of peach leaf extract, and 0.2 parts of birch polypore extract;

[0043] The preparation method is the same as in Example 1, and a green feed additive is prepared.

[0044] Example 3: Preparation of green feed additives:

[0045] Raw material composition (parts by weight): 50 parts of glycoterpenoids, 12 parts of peach leaf extract, and 0.5 parts of birch polypore extract;

[0046] The preparation method is the same as in Example 1, and a green feed additive is prepared.

[0047] Comparative Example 1

[0048] Glycoterpenes can be used alone as a green feed additive.

[0049] Comparative Example 2

[0050] The peach leaf extract prepared using the method of Example 1 alone was used as a green feed additive.

[0051] Comparative Example 3

[0052] The birch polypore extract prepared using the method of Example 1 alone was used as a green feed additive.

[0053] Experimental Example 1: Effects of Green Feed Additives on Broiler Growth Performance

[0054] Experimental Methods: 750 one-day-old broiler chickens, purchased from Dezhou Yuxiyuan Livestock Co., Ltd., were randomly divided into 5 groups, with 3 replicates per group and 50 broiler chickens per replicate. The basal diet for each group was formulated according to the "National Standard for Chicken Feeding—Nutritional Requirements for Broiler Chickens" (ZBB43005-86), and routine feeding and management were implemented. The experiment lasted from 1 day to 40 days of age, and the feeding conditions for each group are as follows:

[0055] T1 treatment group: 1-40 days old were fed a basic diet and the green feed additive prepared in Example 2. For 1-20 days old, 500 mg of green feed additive was added to every 1 kg of basic diet; for 21-40 days old, 1000 mg of green feed additive was added to every 1 kg of basic diet.

[0056] T2 treatment group: 1-40 days old were fed a basal diet and glycosides of Comparative Example 1. From 1-20 days old, 500 mg of glycosides was added per 1 kg of basal diet; from 21-40 days old, 1000 mg of glycosides was added per 1 kg of basal diet.

[0057] T3 treatment group: 1-40 days old were fed a basal diet and peach leaf extract of Comparative Example 2. From 1-20 days old, 500 mg of peach leaf extract was added to every 1 kg of basal diet; from 21-40 days old, 1000 mg of peach leaf extract was added to every 1 kg of basal diet.

[0058] T4 treatment group: 1-40 days old were fed a basal diet and birch polypore extract of Comparative Example 3. From 1-20 days old, 500 mg of birch polypore extract was added to every 1 kg of basal diet; from 21-40 days old, 1000 mg of birch polypore extract was added to every 1 kg of basal diet.

[0059] Control group: fed basic feed from 1 to 40 days old.

[0060] The conservation measures and feeding management are exactly the same for all groups.

[0061] Record the weight changes of broiler chickens: Weigh each surviving broiler chicken at 7, 14, 28 and 40 days of age, and take the average value. Weighing time: half an hour before morning feeding.

[0062] Record the feed intake of each chicken, calculate the survival rate of each treatment group at 40 days of age, and calculate the feed conversion ratio based on the average daily feed intake and average daily weight gain.

[0063] Survival rate (%) = Number of remaining chickens / Original number of chickens × 100%;

[0064] Feed conversion ratio (%) = Average daily feed intake / Average daily weight gain × 100%

[0065] Experimental results:

[0066] Table 1. Changes in broiler chicken weight

[0067]

[0068]

[0069] Table 2 Effects of different feed additives on broiler growth performance

[0070] Group Survival rate (%) Material weight ratio (%) T1 processing group 98.7(148 / 150) 148 T2 processing group 96(144 / 150) 180 T3 processing group 94.7(142 / 150) 191 T4 processing group 95.3(143 / 150) 185 control group 93.3(140 / 150) 202

[0071] As shown in Tables 1 and 2, the compounding of glycosides, peach leaf extract and birch polypore extract in this invention has a significant effect on weight gain in broilers. The addition of the green feed additive of this invention to the basic feed reduced the feed weight ratio by 30.19% compared with the control group, which significantly improved its growth performance.

[0072] Experiment Example 2: Effects of green feed additives on avian influenza and Newcastle disease immunity in broilers

[0073] Test method:

[0074] Two thousand healthy and vigorous 7-day-old broiler chickens were purchased from Dezhou Yuxiyuan Livestock Co., Ltd., and randomly divided into 5 groups, with 4 replicates per group and 100 broiler chickens per replicate. The basal diet for each group was formulated according to the "National Standard for Chicken Feeding - Nutritional Requirements for Broiler Chickens" (ZBB43005-86) and was managed using conventional feeding methods.

[0075] The experiment lasted from 1 day to 40 days of age, and the feeding conditions for each group were as follows:

[0076] Experimental Group 1: 1-40 days old, fed with basal feed and the green feed additive prepared in Example 2, with 1000 mg of green feed additive added per 1 kg of basal diet; Experimental Group 2: 1-40 days old, fed with basal feed and glycosides from Comparative Example 1, with 1000 mg of glycosides added per 1 kg of basal diet; Experimental Group 3: 1-40 days old, fed with basal feed and peach leaf extract from Comparative Example 2, with 1000 mg of peach leaf extract added per 1 kg of basal diet; Experimental Group 4: 1-40 days old, fed with basal feed and birch polypore extract from Comparative Example 3, with 1000 mg of birch polypore extract added per 1 kg of basal diet; Control Group: 1-40 days old, fed with basal feed.

[0077] Immunization trials for ND-HI avian influenza and Newcastle disease were conducted in accordance with the literature "Immunization effect of bivalent inactivated vaccine against Newcastle disease and avian influenza (H9N2 HP strain) in commercial laying hens and commercial broilers" (Heilongjiang Animal Husbandry and Veterinary Medicine, 2017).

[0078] All chickens in Experiments 1-4 were immunized at 7 days of age with a bivalent inactivated vaccine against Newcastle disease (ND) and avian influenza (H9 subtype) (Qingdao Yibang Biotechnology Co., Ltd., 230150125), administered via subcutaneous injection in the neck at a dose of 0.25 mL per chicken. The control group was not immunized and served as a blank control. Blood samples were randomly selected from each of Experiments 1-4 and the control group for antibody monitoring at 7 days of age and on days 7, 14, 21, 28, and 35 post-immunization (the geometric mean of the antibody results from the 10 chickens was used). For the remaining chickens in Experiments 1-4 and the control group, 10 chickens were randomly selected on days 14 and 21 post-immunization for avian influenza antibody testing. Following blood collection, an avian influenza virus challenge protection test was conducted.

[0079] Antibody detection method: Antibody detection follows the 2000 edition of the Regulations for Veterinary Biological Products of the People's Republic of China, using the erythrocyte agglutination inhibition test method. The serum to be tested is serially diluted 2-fold with PBS, and hemagglutinin antigen solution containing 4 HAU is added. PBS and 4 HAU hemagglutinin antigen solutions are included as controls. After thorough shaking, the solution is incubated at room temperature for at least 20 min or at 2–8°C for 40–60 min. Then, 1% chicken erythrocyte suspension is added, and the solution is incubated at room temperature for 20–40 min or at 2–8°C for 40–60 min. The result is determined when the erythrocytes in the control wells show a distinct button-like appearance. The highest serum dilution that completely inhibits erythrocyte agglutination is used as the endpoint.

[0080] Table 3 Newcastle disease antibody levels Nlog2 (N is the reading in the table)

[0081] Group Pre-immunization 7 days after immunization 14 days after immunization 21 days after immunization 28 days after immunization 35 days after immunization Experiment 1 group 4.9 3.8 6.2 8.7 9.5 9.8 Experimental Group 2 4.8 3.7 4.2 6.3 6.5 6.6 Experiment 3 groups 5.0 3.7 4.1 5.6 6.3 6.4 Experiment 4 groups 4.9 3.6 5.2 6.1 6.2 6.3 control group 5.0 3.3 1.8 0.9 0.2 0

[0082] Table 4. Avian influenza antibody levels Nlog2 (N is the reading in the table)

[0083] Group Pre-immunization 7 days after immunization 14 days after immunization 21 days after immunization 28 days after immunization 35 days after immunization Experiment 1 group 3.2 1.9 6.3 7.8 8.9 9.0 Experimental Group 2 3.1 1.8 3.8 4.7 5.9 6.5 Experiment 3 groups 3.3 1.5 2.9 4.2 6.0 6.1 Experiment 4 groups 3.0 1.2 3.2 4.3 6.3 6.4 control group 3.2 0.9 0.6 0.2 0 0

[0084] As shown in Tables 3 and 4, before vaccination, there were no significant differences in antibody levels among the groups compared to the control group (P>0.05). However, 14 days after immunization, the antibody levels in the feed supplemented with the green feed additive were significantly higher than those in the control group (P<0.05). This demonstrates that the green feed additive of this invention can enhance immunity, promote antibody secretion, accelerate immune cell proliferation, and enhance antigen-antibody levels.

[0085] The above description is merely a preferred embodiment of this application and is not intended to limit this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the protection scope of this application.

Claims

1. A green feed additive, characterized in that, The green feed additive comprises the following components in parts by mass: 40 parts of sugar terpenoid, 10 parts of peach leaf extract, and 0.2 parts of birch mushroom extract; The peach leaf extract is obtained by the following method: 80-95% ethanol by volume is added to peach leaf powder, and ultrasonic extraction is performed for 2-4 times, followed by filtration to obtain an extract; the extract is concentrated and dried to obtain the peach leaf extract; The birch mushroom extract is obtained by the following method: (1) The birch mushroom is crushed and sieved, and water is added as a fermentation medium; (2) A mixed seed liquid is inoculated into the fermentation medium, the inoculation amount of the mixed seed liquid is 5-10% of the volume of the fermentation medium, the mixed seed liquid is mixed by Brevibacillus parvus, Candida utilis, and Bacillus coagulans; constant temperature fermentation is performed at 30-35°C for 60-72h, followed by filtration to obtain a fermentation liquid; the fermentation liquid is concentrated and dried to obtain the birch mushroom extract.

2. The green feed supplement according to claim 1, characterized in that, The solid-liquid ratio of the peach leaf powder and ethanol is 1g:(20-30)ml; the ultrasonic extraction time is 10-15min, and the ultrasonic power is 150-200W.

3. The green feed supplement according to claim 1, characterized in that, In step (1), the mass ratio of birch mushroom to water is 1:15-20.

4. The green feed supplement according to claim 1, characterized in that, In step (2), the concentration of B. laterosporus in the mixed seed solution is 1.0 x 10 8 ~1.0 x 10 9 cfu / mL, the concentration of C. utilis is 1.0 x 10 7 ~1.0 x 10 8 cfu / mL, and the concentration of B. coagulans is 1.0 x 10 7 ~1.0 x 10 8 cfu / mL.

5. The method for preparing the green feed additive according to any one of claims 1 to 4, characterized in that, The method comprises the following steps: The sugar terpenoid, peach leaf extract, and birch mushroom extract are weighed according to the mass parts, mixed uniformly, and a green feed additive is obtained. The green feed additive comprises the following components in parts by mass: 40 parts of sugar terpenoid, 10 parts of peach leaf extract, and 0.2 parts of birch mushroom extract; The peach leaf extract is obtained by the following method: 80-95% ethanol by volume is added to peach leaf powder, and ultrasonic extraction is performed for 2-4 times, followed by filtration to obtain an extract; the extract is concentrated and dried to obtain the peach leaf extract; The birch mushroom extract is obtained by the following method: (1) The birch mushroom is crushed and sieved, and water is added as a fermentation medium; (2) A mixed seed liquid is inoculated into the fermentation medium, the inoculation amount of the mixed seed liquid is 5-10% of the volume of the fermentation medium, the mixed seed liquid is mixed by Brevibacillus parvus, Candida utilis, and Bacillus coagulans; constant temperature fermentation is performed at 30-35°C for 60-72h, followed by filtration to obtain a fermentation liquid; the fermentation liquid is concentrated and dried to obtain the birch mushroom extract. The solid-liquid ratio of the peach leaf powder and ethanol is 1g:(20-30)ml; the ultrasonic extraction time is 10-15min, and the ultrasonic power is 150-200W. In step (1), the mass ratio of birch mushroom to water is 1:15-20. The method comprises the following steps: The sugar terpenoid, peach leaf extract, and birch mushroom extract are weighed according to the mass parts, mixed uniformly, and a green feed additive is obtained.

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