A medicinal and edible composition, a preparation method and application thereof

Abstract

The present application belongs to the field of feed and traditional Chinese veterinary medicine, and particularly relates to a kind of medicinal and edible composition and its preparation method and application.The present application provides a kind of medicinal and edible composition, which is made of liquorice, gold buckwheat and thistle seed.The gold buckwheat, liquorice and thistle seed in the medicinal and edible composition of the present application can achieve synergistic effect, and can achieve the effects of improving the growth performance of pigs, reducing the feed-meat ratio, improving the stress resistance of pigs, improving the liver performance of pigs, improving the immunity of pigs, and improving the intestinal flora of pigs.

Description

Technical Field

[0001] The present invention belongs to the fields of feed and traditional Chinese veterinary medicine, and particularly relates to a medicine-food homologous composition, a preparation method thereof, and an application thereof. Background Art

[0002] Medicine-food homologous plants originate from nature and have advantages such as being natural, having low toxicity, and having few residues. At present, studies have found that some medicine-food homologous plants can improve the production performance of piglets and enhance immunity, and have good health care effects on piglets. For example, astragalus membranaceus, wolfberry, hawthorn, etc., which are widely used clinically, all have anti-inflammatory and antioxidant effects. Adding the formula of Jingfang Baidu powder to the diet can significantly improve the growth rate and weight gain effect of pigs. Current studies have found that medicine-food homologous plants, as feed additives, can promote the growth and development of weaned piglets and lactating sows.

[0003] Currently used plant or plant extract type additives are all substances that use single or mixed plants and retain the original biological activities and natural factors of the plants, mainly including flavonoids, polyphenols, polysaccharides, alkaloids, organic acids, volatile oils, and saponins. Most of these bioactive components are secondary metabolites in the process of plant growth and development and have biological functions such as promoting growth, antioxidant, antibacterial, antiviral, and enhancing immunity. Current plant or extract type additives often increase animal feed intake, stimulate the secretion of growth-related hormones, and promote bone tissue growth through single or synergistic effects between active components, so as to achieve the effect of promoting animal growth. At present, there are few studies on using medicine-food homologous plant formulas as feed additives to improve pig feed utilization rate, immune function, and antioxidant capacity. Therefore, developing new plant feed additives with rich functions and capable of replacing antibiotics is an urgent problem to be solved at present. Summary of the Invention

[0004] In order to solve the above problems, the present invention provides a medicine-food homologous composition, which is made of licorice, fagopyrum dibotrys, and arctium lappa.

[0005] A medicine-food homologous composition is made of 10-30 parts of licorice, 20-40 parts of fagopyrum dibotrys, and 30-70 parts of arctium lappa.

[0006] Further, the medicine-food homologous composition is made of 10-20 parts of licorice, 20-30 parts of fagopyrum dibotrys, and 40-70 parts of arctium lappa.

[0007] Further, the medicine-food homologous composition is made of 10-15 parts of licorice, 25-30 parts of fagopyrum dibotrys, and 55-70 parts of arctium lappa.

[0008] Further, the medicine-food homologous composition is made of 10 parts of licorice, 30 parts of fagopyrum dibotrys, and 60 parts of arctium lappa.

[0009] The present invention also provides a method for preparing the aforementioned food-medicine homology composition, specifically, by weighing licorice, buckwheat, and burdock seeds according to the mass ratio, mixing them, drying them to a moisture content of ≤10%, and then pulverizing them into powder with a mesh size of ≥300 to obtain the final product.

[0010] The present invention also provides a feed additive containing the aforementioned food-medicine homology composition.

[0011] The present invention also provides a feed containing the aforementioned food-medicine homology composition.

[0012] The present invention also provides the application of the aforementioned food-medicine homology composition in the preparation of feed or medicine that improves pig growth performance, stress resistance, liver performance, immunity, and intestinal flora.

[0013] Furthermore, the amount of the medicinal and edible homology composition added to the feed is 0.1% to 1%.

[0014] Furthermore, the amount of the medicinal and edible homology composition added to the feed is 0.3%.

[0015] The advantages of this invention over the prior art are as follows:

[0016] In view of the fact that currently used plant or plant extract additives rely solely on secondary metabolites such as flavonoids, polyphenols, polysaccharides, and alkaloids to achieve biological functions such as growth promotion, antioxidation, antibacterial, antiviral and immune enhancement, this invention provides a medicinal and edible plant composition suitable for low-protein diets for pigs.

[0017] The combination of buckwheat, licorice, and burdock seed has been tested and found to have a synergistic effect, which can improve the growth performance of pigs, reduce the feed conversion ratio, improve pig stress, improve pig liver performance, enhance pig immunity, and improve pig intestinal flora. Attached Figure Description

[0018] Figure 1 The total ion current chromatogram of the mixed reference standard shows that peak 1 is chlorogenic acid, peak 2 is naringin, peak 3 is arctiin, and peak 4 is epicatechin gallate.

[0019] Figure 2 For the detection of serum liver function biochemical indicators (n=5).

[0020] Figure 3 The effect of plant-based feed additives containing medicinal herbs on antioxidant enzyme levels in fattening pigs (n=5).

[0021] Figure 4 Serum concentrations of cytokines IL-6 and TNF-α (n=5)

[0022] Figure 5For PCoA analysis based on the Bray-Curtis distance matrix of OTUs, each point in the figure represents one sample, samples in the same group are represented by the same color, and the colored area represents the confidence interval. The same labeled symbol represents four samples in the same group. Permanova, p = 0.001.

[0023] Figure 6 The study aimed to identify the gut microbiota structure characteristics of pigs after feeding with stress-resistant feed, including: a) Venn diagram based on OTUs; b) relative abundance analysis of dominant gut microbiota genera; c) cluster analysis of gut microbiota genera in each sample; d) heatmap of species abundance at the genus level in each group and sample; and e) α-diversity analysis.

[0024] Figure 7 To identify biomarkers in different groups for LEfSe analysis, the LDA value distribution bar chart mainly shows species with LDA values ​​greater than the preset values ​​(generally, the less strict screening threshold is set to 2; the more strict screening threshold is set to 4), i.e., biomarkers with statistical differences. The color of the bar chart represents the respective group, and the length of the bar represents the LDA value, i.e., the degree of influence of species with significant differences between different groups. In the clade diagram, the circles radiating outward represent the taxonomic levels from kingdom (single circle) to genus (or species). Each small circle at a different taxonomic level represents a taxonomy at that level, and the diameter of the small circle is proportional to the relative abundance of the species. Species with no significant differences are uniformly colored yellow, while species with significant differences are colored according to their group. Different colors indicate microbial groups that play an important role in their respective groups. The species names corresponding to the biomarkers are shown on the right, with letter codes corresponding to those in the figure.

[0025] Figure 8 This is for KEGG function prediction based on gut microbiota 16S. Detailed Implementation

[0026] The embodiments of the present invention will be described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are for illustrative purposes only and should not be considered as limiting the scope of the invention. Unless otherwise specified in the examples, conventional conditions or conditions recommended by the manufacturer are followed. Reagents or instruments whose manufacturers are not specified are all commercially available products.

[0027] The raw materials, buckwheat, licorice, and burdock seeds, used in the following examples are selected from the "Catalogue of Plants with Both Medicinal and Feed Properties" (2022).

[0028] The main chemical components of buckwheat, licorice, and burdock seed, namely epicatechin gallate, chlorogenic acid, and arctiin, were used as targets for detection.

[0029] Quality testing method: The target components were determined by high performance liquid chromatography (HPLC). For details, please refer to the following: Thermo Hypersil Gold-C18 (4.6 mm × 250 mm, 5 μm) column was used, the mobile phase was acetonitrile-0.1% phosphoric acid solution (9:91), the flow rate was 1.0 mL·min⁻¹, the column temperature was 30℃, the detection wavelength was 280 nm, and the injection volume was 10 μL.

[0030] The test results of the food and medicine plant additives are shown in Table 1. Their quality standards meet the quality standards of each single Chinese medicine in the Chinese Veterinary Pharmacopoeia (2020 edition).

[0031] Table 1 Minimum Content of Effective Components in Medicinal and Edible Plants

[0032] Element CAS number Minimum detectable concentration (μg / g) Epicatechin gallate 1257-08-5 120.00 chlorogenic acid 327-97-9 950.00 Arctiin 20362-31-6 15.00

[0033] Example 1: Preparation of a food-medicine homology composition

[0034] (1) Mix 10 parts of licorice, 30 parts of buckwheat and 60 parts of burdock according to the mass ratio, dry them to make the moisture content ≤10%, and then pulverize them into powder with a mesh size ≥300.

[0035] (2) Mix 10 parts of licorice, 20 parts of buckwheat and 70 parts of burdock according to the mass ratio, dry them to make the moisture content ≤10%, and then pulverize them into powder with a mesh size ≥300.

[0036] (3) Mix 15 parts of licorice, 30 parts of buckwheat and 55 parts of burdock according to the mass ratio, dry them to make the moisture content ≤10%, and then pulverize them into powder with a mesh size ≥300.

[0037] (4) Mix 20 parts of licorice, 30 parts of buckwheat and 50 parts of burdock according to the mass ratio, dry them to make the moisture content ≤10%, and then pulverize them into powder with a mesh size ≥300.

[0038] (5) Mix 20 parts of licorice, 20 parts of buckwheat and 60 parts of burdock according to the mass ratio, dry them to make the moisture content ≤10%, and then pulverize them into powder with a mesh size ≥300.

[0039] (6) Mix 20 parts of licorice, 25 parts of buckwheat and 40 parts of burdock according to the mass ratio, dry them to make the moisture content ≤10%, and then pulverize them into powder with a mesh size ≥300.

[0040] (7) Mix 30 parts of licorice, 40 parts of buckwheat and 30 parts of burdock according to the mass ratio, dry them to make the moisture content ≤10%, and then pulverize them into powder with a mesh size ≥300.

[0041] When using, add 0.3% of the medicinal and edible plant formula to the low-protein complete diet or other complete diet of pigs according to the mass ratio, mix thoroughly and feed to pigs. There is no limit to the feeding time.

[0042] Example 2: Detection of representative components in anti-stress feed

[0043] According to "Preparation of Food Additives from Medicinal and Edible Plants in Example 1", "(1) Mix 10 parts of licorice, 30 parts of buckwheat and 60 parts of burdock seed according to the mass ratio, dry them to make the moisture content ≤10%, and prepare them into powder with a mesh size ≥300" and name it Formula A.

[0044] Formula A, consisting of medicinal and edible plants, was added to the piglet basal feed at mass ratios of 0.1% and 0.3%, respectively, and named experimental groups A1 and A2. The main chemical components of buckwheat, licorice, and burdock seed—epicoetin gallate (1257-08-5), chlorogenic acid (327-97-9), and arctiin (20362-31-6)—were detected using high-performance liquid chromatography (HPLC). The basal feed was based on a low-protein diet; the low-protein basal feed formula is shown in Table 2, and its nutrient composition ratio was analyzed, as shown in Table 3.

[0045] Table 2 Low-protein base feed formulation

[0046]

[0047]

[0048] Table 3. Nutritional component percentages in low-protein base feed formulations

[0049]

[0050]

[0051]

[0052] The testing process is as follows:

[0053] (1) Sample pretreatment

[0054] Weigh approximately 0.2g of each feed sample, add 1mL of methanol extract, homogenize in an ice bath (for liquid samples, dilute twice with the extract and mix well), sonicate in an ice-water bath for 60min, centrifuge at 8000g for 10min, collect the supernatant, filter it through a syringe filter and then test.

[0055] (2) Chromatographic analysis conditions

[0056] Thermo U3000 high performance liquid chromatograph, Thermo C18 reversed-phase column (250mm*4.6mm, 5μm), mobile phase acetonitrile-0.1% phosphoric acid solution (9:91), flow rate 0.8mL·min⁻¹, column temperature 30℃, detection wavelength 280nm, injection volume 10μL.

[0057] (3) Standard curve and standard sample spectrum

[0058] Different masses of chlorogenic acid, naringin, arctiin, and epicatechin gallate were weighed and placed in 25L brown volumetric flasks. 70% methanol was added and the mixture was brought to volume. The mixture was shaken well to prepare the reference standards. The total ion chromatogram of the reference standards is shown in [Figure number missing]. Figure 1 .

[0059] Accurately measure an appropriate amount of the mixed reference standard stock solution, dilute it with 70% methanol solution to obtain a series of mixed reference standards with different mass concentrations, and then determine them. Plot the mass concentration C (ng / mL) as the abscissa and the peak area A as the ordinate, and calculate the linear regression equations of the four components. The results are shown in Table 4.

[0060] Table 4. Results of linear relationships (n=4)

[0061] Indicator Name Standard curve <![CDATA[R 2 ]]> Retention time (min) chlorogenic acid y = 0.287x - 0.0061 0.9999 5.170 Naringin y = 0.3931x - 0.0285 0.9999 12.877 Arctiin y = 0.3818x - 0.0047 0.9997 19.063 Epicatechin gallate y = 0.1174x - 0.0149 0.9997 28.662

[0062] According to the mass ratio, the medicinal and edible plant formulation A was added to the basal feed of piglets at 0.1% (A1) and 0.3% (A2). The contents of the main chemical components of buckwheat, licorice and burdock seed, namely epicatechin gallate (1257-08-5), chlorogenic acid (327-97-9) and arctiin (20362-31-6), in the basal feed group (N), A1 and A2 groups were detected by high performance liquid chromatography. The detection results are shown in Table 5.

[0063] Table 5. Detection values ​​of representative components in anti-stress pig feed

[0064]

[0065] Example 3: Growth performance and stress resistance test

[0066] According to "Preparation of Food Additives from Medicinal and Edible Plants in Example 1", "(1) Mix 10 parts of licorice, 30 parts of buckwheat and 60 parts of burdock seed according to the mass ratio, dry them to make the moisture content ≤10%, and prepare them into powder with a mesh size ≥300" and name it Formula A.

[0067] The medicinal and edible plant additive (Formula A) designed in this invention was used in combination with a low-protein diet to feed Landrace growing-finishing pigs that had not been vaccinated against various diseases. The pigs were of equal sex. The experiment was divided into five groups, with 12 pigs in each group, set up with three replicates of four pigs per replicate, and fed in separate pens. Formula A was added to the low-protein basal feed at 0.1% and 0.3% to prepare pig feed, named experimental groups A1 and A2, respectively. Simultaneously, commercially available functional feed additives, rosemary extract and seaweed polysaccharide extract, were used as control groups P1 and P2, respectively, and a control group N for the low-protein basal feed was also set up (see Table 6). Before the experiment, the pigpens, water troughs, and feed troughs were disinfected with 3% caustic soda. The experimental fattening pigs were dewormed and fed for two weeks until they were confirmed to be healthy and disease-free before the experiment began. Throughout the experiment, all fattening pigs were kept in pens with free access to water. Before feeding, the animals were fasted for 24 hours but allowed to drink water. Before being separated into pens, their average weight was measured to be 19.85 ± 4.23 kg. The feeding period was 45 days. After the feeding period, the animals were fasted for 24 hours but allowed to drink water, and were weighed again. During this period, the total amount of feed used by each group was recorded, and the daily weight gain and feed conversion ratio were calculated.

[0068] Table 6 Experimental Groups

[0069]

[0070] The base feed is based on low-protein feed. The formula of the low-protein base feed is shown in Tables 2 and 3 of “Representative Component Detection of Anti-Stress Feed in Example 2”.

[0071] 3.1 Evaluation of growth-promoting effects

[0072] Feed twice daily, morning and evening, weighing the feed each time. Allow free access to food and water, and use a slatted floor. The feeding period is 45 days. On the last day of feeding, withhold food but allow free access to water for 24 hours, and weigh the pigs again. Weigh 1-2 pigs per replicate, and weigh 5 pigs per group, taking the average weight. The weight gain during the feeding period ranged from 32.38 to 33.62 kg, with a daily weight gain of 0.720 to 0.747 kg and a feed conversion ratio of 2.255 to 2.575. Comprehensive evaluation revealed that A1 (0.1% of formula A) and A2 (0.3% of formula A) both showed good effects, with weight gains of 32.97 and 33.38 kg, daily weight gains of 0.733 and 0.742 kg, and feed conversion ratios of 2.257 and 2.305, respectively. The weight gains during the feeding period for the commercial functional additives P1 (rosemary extract added group) and P2 (seaweed polysaccharide extract added group) were 33.35 and 33.62 kg, with daily weight gains of 0.741 and 0.747 kg, and feed conversion ratios of 2.436 and 2.255, respectively (see Table 7). Both A1 and A2 were superior to the blank control group.

[0073] Table 7 Results of the feeding trial of stress-resistant pig feed

[0074]

[0075]

[0076] 3.2 Disease incidence during the feeding period

[0077] During the 45-day feeding period, the incidence of diarrhea, vomiting, rectal prolapse, other abnormalities (increased eye discharge, cyanosis of ear tips, necrosis, lethargy for 1-3 days, etc.), and death in each group of pigs were recorded. Pigs exhibiting diarrhea, vomiting, or rectal prolapse were not treated; they were isolated and fed until recovery, and then returned to their original pens. This was counted as one outbreak. If the symptoms recurred 5 days after recovery, it was counted as another outbreak. Observation showed that, except for groups A2 and P2, all other groups had 1-4 outbreaks, with an incidence rate of 8.3%-33.3%. The incidence rate in group N was 25.0% (see Table 8). Both A1 and A2 were superior to the blank control group.

[0078] Table 8. Disease incidence during the feeding period

[0079]

[0080]

[0081] Example 4 Liver Function Test

[0082] After the feeding period, pigs in the blank control group (N) and the group with 0.3% added A group formula (A2) in "Example 2 Feeding Effect" were fasted for 24 hours but allowed free access to water. Blood was collected via the anterior vena cava, and serum was separated. Samples were collected from 1-2 pigs per replicate, with 5 pigs collected from each group. Serum gamma-glutamyl transferase (GGT), total bilirubin (TBIL), direct bilirubin (DBIL), indirect bilirubin (IBIL), and triglyceride (TG) levels were measured. These indicators reflect the state of liver parenchymal cells; lower values ​​indicate less damage and better function of liver parenchymal cells. The serum samples from the 5 pigs in each group were analyzed using box plots. The results showed significant individual differences in these indicators among the groups, but all were within the normal range. There were no significant differences between the two groups (P>0.05), indicating that the addition of the A group's herbal additive was not toxic to the liver, and the values ​​in group A2 were lower than those in the blank control group (see...). Figure 2 The results indicate that adding the A-group medicinal and feed homologous plant additives can reduce the damage to the liver parenchyma in pigs.

[0083] Example 5 Antioxidant and Immunological Performance Tests

[0084] After the feeding period of pigs in the blank control group (N) and the group with 0.3% added A formula (A2) in "Feeding Effect of Example 2" was completed, they were fasted for 24 hours but allowed to drink water. Blood was collected through the anterior vena cava and serum was separated. Samples were collected from 1-2 pigs in each replicate, and samples were collected from 5 pigs in each group for serum antioxidant stress index and immune cytokine detection.

[0085] 5.1 Detection of serum antioxidant stress markers

[0086] Oxidative reactions such as respiration and metabolism in organisms generate reactive free radicals. Under normal conditions, free radicals have a stable scavenging system, maintaining a low concentration balance in the body. Oxidative stress refers to the process of oxidative damage caused by an imbalance between the production and scavenging of oxygen free radicals in the body or cells, leading to the accumulation of reactive oxygen species (ROS) and reactive nitrogen species (RNS) in the body or cells. Detecting antioxidant indicators can assess the quality of a feed additive. In studies, the antioxidant capacity of an additive can be evaluated by measuring the levels of total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-Px), catalase (CAT), superoxide dismutase (SOD), and malondialdehyde (MDA) in serum. Total antioxidant capacity (T-AOC) refers to the total antioxidant level composed of various antioxidant substances and enzymes, such as antioxidant enzymes, vitamin C, vitamin E, and carotene. To protect cells and the body from oxidative stress damage caused by reactive oxygen free radicals, total antioxidant capacity can be used to evaluate the antioxidant capacity of bioactive substances. Glutathione peroxidase (GSH-Px) is an important peroxide-degrading enzyme widely present in the body. The active center of GSH-Px is selenocysteine, and its activity reflects the body's selenium level. Selenium is a component of the GSH-Px enzyme system; it catalyzes the conversion of GSH to GSSG, reducing toxic peroxides to non-toxic hydroxyl compounds, thereby protecting the structure and function of cell membranes from interference and damage by peroxides. Catalase (CAT) is an enzyme that catalyzes the decomposition of hydrogen peroxide into oxygen and water, and is present in the peroxisome of cells. Catalase is a marker enzyme of peroxisomes, accounting for approximately 40% of the total peroxisome enzymes. Catalase is present in all known animal tissues, particularly in high concentrations in the liver. Superoxide dismutase (SOD) is an antioxidant metalloenzyme found in organisms that catalyzes the dismutation of superoxide anion free radicals into oxygen and hydrogen peroxide. It plays a crucial role in the body's oxidation-antioxidant balance and is closely related to the occurrence and development of many diseases. Malondialdehyde (MDA) content is an important parameter reflecting the body's antioxidant potential, indicating the rate and intensity of lipid peroxidation and indirectly reflecting the degree of tissue peroxidation damage. In vivo, free radicals act on lipids to cause peroxidation, with malondialdehyde as the final oxidation product. This can lead to cross-linking and polymerization of biomolecules such as proteins and nucleic acids, and is cytotoxic.

[0087] Formula A was added at a concentration of 0.3% to a low-protein basal feed to prepare pig feed. A control group (N) was set up to feed the pigs for 45 days. Serum samples were collected from 5 pigs in each group for testing serum antioxidant indicators. The results showed that the serum T-AOC and T-SOD levels in group A2 (with 0.3% of formula A added) were higher than those in the blank control group (N group), and CAT and GSH-Px were significantly higher in group A2 than in the blank control group (N group). The MDA antioxidant index was also better in group A2 than in the blank control group (N group). (See figure) Figure 3 This indicates that adding 0.3% of Group A can protect cells such as the liver from oxidative stress damage caused by reactive oxygen free radicals and promote the body's health.

[0088] 5.2 Cytokine Detection

[0089] Cytokines (CK) are small polypeptides or glycoproteins synthesized and secreted by various tissue cells (mainly immune cells). Cytokines mediate cell-cell interactions and have a variety of biological functions, such as regulating cell growth, differentiation and maturation, maintaining function, regulating immune responses, participating in inflammatory responses, wound healing, and tumor growth and decline.

[0090] Interleukin-6 (IL-6) is a cytokine belonging to the chemokine family and is one of the most typical cytokines associated with inflammation. IL-6 is a pleiotropic cytokine that can promote the proliferation and differentiation of various cell types and plays an important role in host defense by regulating immune and inflammatory responses. Tumor necrosis factor-alpha (TNF-α) is a pleiotropic cellular molecule that plays a central role in inflammation, apoptosis, and immune system development. Primarily produced by macrophages and monocytes, it can synergistically regulate the production of other cytokines, cell survival, and death to coordinate tissue homeostasis. TNF-α has diverse biological functions and a complex mechanism of action. In healthy individuals, TNF-α can enhance resistance to infection by activating neutrophils and platelets, enhancing the killing ability of macrophages / NK cells, and stimulating the immune system. Higher concentrations of IL-6 and TNF-α are beneficial in healthy individuals.

[0091] The test results showed that the levels of IL-6 and TNF-α were both within the normal physiological concentration range. The IL-6 level in group A2 was significantly higher than that in the blank control group (group N), ranging from 120 to 180 pg / mL. The concentration of TNF-α in group A2 was slightly higher than that in the blank control group (group N). Figure 4 .

[0092] Example 6: Analysis of gut microbiota after feeding

[0093] The low-protein feed formulation, grouping, and feeding method are described in Example 2. Additionally, a C2 group was set up to replace the A1 group. Group C2: Formula C was added at 0.3%. Formula C: 30 parts licorice, 40 parts bitter orange peel, and 30 parts buckwheat were mixed according to the mass ratio, and the production method is described in Example 1. After feeding, fecal microbiota analysis was performed on the blank control group (N), the rosemary extract control group (P1), the seaweed extract control group (P2), the A2 group (with 0.3% of Formula A added), and the C2 group (with 0.3% of Formula C (a mixture of 30 parts licorice, 40 parts bitter orange peel, and 30 parts buckwheat, according to the mass ratio)). Rosemary extract contains various functional chemical substances such as rosemary essential oil, carrageenan, oleanolic acid, ursolic acid, and rosmarinic acid. After feeding, these substances are mostly absorbed into the bloodstream, thereby promoting animal growth, immunity, antioxidant effects, and meat quality. Seaweed and seaweed extracts primarily promote health, immune regulation, and production performance by increasing gut microbiota and promoting the synthesis of short-chain fatty acids. After the feeding trial, rectal feces were collected after a 12-hour fast and frozen in liquid nitrogen for later use. 16S rDNA amplicon sequencing analysis of the gut microbiota was performed by Beijing Aovisen Gene Technology Co., Ltd. using the NovaSeq PE250 protocol. After read splicing and filtering, operational taxonomic units (OTUs) clustering, species annotation, abundance analysis, alpha diversity analysis, and beta diversity analysis were conducted.

[0094] 6.1 Analysis of differences in gut microbiota grouping

[0095] Adding traditional Chinese medicine to anti-stress pig feed may lead to changes in gut microbiota, which may produce anti-stress effects. The gut microbiota structure was studied using 16S rDNA amplicon sequencing technology. First, principal coordinates analysis (PCoA) based on the Bray-Curtis distance matrix was performed to reflect the β-diversity of different groups. (PCoA diagram follows.) Figure 5 The results showed that there were significant differences in gut microbiota among the groups (PerMANOVA, P = 0.001), indicating that each group could form its own characteristic microbiota.

[0096] 6.2 Characteristics and Changes of Gut Microbiota

[0097] Analysis of the composition and structure of the gut microbiota revealed that the number of OTUs in groups N, P1, P2, A2, and C2 were 4139, 3959, 3960, 3678, and 3873, respectively. A total of 2670 OTUs were shared across the five groups, but only 577 OTUs were shared among individuals. Figure 6 -a).

[0098] Analysis of the dominant bacterial genera in each group revealed that group A2 and seaweed extract (P2) had similar mechanisms of action, increasing the relative abundance of beneficial bacteria in the pig gut, including *Prevotella*, *Prevotellaceae NK3B31* group, *Muribaculaceae*, *Roseburia*, *Clostridium sensu stricto* 1, *Prevotellaceae UCG-003*, *Clostridium sensu stricto* 6, *Faecalibacterium*, *Lachnospiraceae NK4A136* group, *Lactobacillus*, *Alloprevotella*, and *Agathobacter*. The abundances of these beneficial bacteria were 60.0% and 57.3%, respectively, significantly higher than those in group N (50.8%), group P1 (39.9%), and the control group C2 (48.0%). (See [link to relevant documentation]). Figure 6 -b. Cluster analysis was performed on the top 15 genera in terms of relative abundance. Figure 6 -c), it was found that at the genus level, A2 and the group that regulates the body through improving gut microbiota by adding seaweed extract (P2 group) clustered on the same branch, and were relatively close to the N group, C2 and P1 group that clustered on the same branch. An evolutionary distance heatmap was drawn for the 20 most abundant bacterial genera. Figure 6-d) shows that Prevotellaceae_NK3B31_group, Prevotella, Streptococcus, Clostridium sensu_stricto_1, Faecalibacterium, Roseburia, Agathobacter (Agathobacter is a Gram-positive anaerobic bacterium whose main products in the culture medium are butyric acid and acetic acid), Lactobacillus, and Succinivibrio were abundant in the A2 group samples. The abundance of *Muribaculaceae*, *Treponema*, *Ruminococcaceae* (UCG-005), and *Prevotellaceae* (UCG-003) was relatively high. In group C2, *Prevotellaceae_NK3B31_group*, *Prevotella*, *Muribaculaceae*, *Lactobacillus*, and *Succinivibrio* were abundant, while *Streptococcus* was less so. The α-diversity index of the gut microbiota can directly reflect the species diversity within the group. Figure 6 The -e option shows differences in the Chao1, Shannon, Simpson, and PD_whole_tree indices among the five groups. The lower Chao1 and PD_whole_tree indices in group A2 indicate that group A2 has fewer bacterial species and closer phylogenetic relationships. It is possible that the higher abundance of probiotics in group A2 inhibits the growth of harmful bacteria.

[0099] 6.3 Differential Microbial Community Analysis

[0100] To further analyze the differences in gut microbiota composition among the groups, LEfSe analysis (Linear Discriminant Analysis Effect Size) was performed to identify biomarkers with significant differences in abundance among the groups. Figure 7The linear discriminant analysis (LDA) value distribution histogram shows that multiple biomarkers exist in each group of gut microbiota. In group N, there are 2 biomarkers with LDA values ​​greater than 4, including the family Muribacaceae and the genus Muribacaceae; in group P1, there are 7 biomarkers with LDA values ​​greater than 4, including the genus Enterococcaceae bacterium RF39, the family Enterococcaceae bacterium RF39, the order Enterococcaceae bacterium RF39, the family Ruminococci (UCG-005), the order Lactobacillales, the class Bacilli, and the phylum Firmicutes; in group P2, there are 8 biomarkers with LDA values ​​greater than 4, including the kingdom Bacteria, the family Streptococcaceae, the genus Streptococcaceae, and the family Prevotellaceae (NK3B31). The groups included Bacteroidetes, Bacteroidetes, Bacteroidetes, and Prevotella; Group A2 contained 8 species with LDA values ​​greater than 4, including Agathobacter, Faecalibacterium, Ruminococci, Rochetomyces, Clostridia, Lachnospiraceae, Lachnospirales, and Prevotella; Group C2 contained 15 species with LDA values ​​greater than 4, including Treponema, Spirogyra, and Prevotella. The bacterial flora belongs to the phylum Spirochaetota, order Spirochaetales, family Spirochaetaceae, class Spirochaetia, family Succinivibrionaceae, phylum Proteobacteria, order Aeromonadales, class Gammaproteobacteria, genus Succinivibrio, species Lactobacillus vaginalis, family Oscillospiraceae, genus Lactobacillus, family Lactobacillaceae, and order Oscillospirale. Among these, group A2 bacteria exhibit a higher concentration of probiotic biomarkers for the synthesis of short-chain fatty acids (SCFAs) such as butyrate, which can induce the production of IL-22 by CD4+ T cells and innate lymphocytes (ILCs) in pigs to maintain intestinal homeostasis and promote overall health.

[0101] PICRUSt2 (2.3.0b0) was used to predict the composition of known microbial gene functions based on bacterial 16S sequences, thereby statistically analyzing functional differences between different samples and groups. KEGG-based level 3 functional predictions indicate that these five groups are involved in carbohydrate metabolism, primarily involving galactose metabolism, starch and sucrose metabolism, and C5-branched dibasic acid metabolism; other glycan degradation and glycosaminoglycan degradation (glycan biosynthesis and metabolism); lipoic acid metabolism; thiamine metabolism; riboflavin metabolism; vitamin B6 metabolism (metabolic metabolism of cofactors and vitamins); biosynthesis of ansamycins; and biosynthesis of vancomycin antibiotics. Differences exist in the abundance of functional genes in the metabolism of antibiotics (terpenoids and polyketides) and drug metabolism-other enzymes (xenobiotics biodegradation and metabolism).

[0102] Comparative studies revealed that the abundance of functional genes related to carbohydrate metabolism in the rectal flora of groups A2 and P2 was significantly higher than that of the control group (N group). Furthermore, group A2 outperformed group P2 in starch and sucrose metabolism and C5 branched-chain dicarboxylic acid metabolism, indicating that the A2 formula can enhance the body's utilization of carbohydrates in feed and improve feed conversion efficiency. However, the abundance of functional genes related to glycosaminoglycan degradation and other sugar degradation in groups A2 and C2 was significantly lower than that in groups N and P2. The abundance of functional genes related to thiamine metabolism, riboflavin metabolism, and vitamin B6 metabolism in the rectal flora of groups A2 and P2 was significantly higher than that in the control group (N group). B vitamins are essential nutrients for body tissues and are crucial for the release of energy from food. They work synergistically to participate in the metabolism of sugars, proteins, and fats in the body, indicating that the A2 formula can enhance the synthesis and metabolism of sugars, proteins, and fats in the body and improve feed conversion efficiency. In lipoic acid metabolism, the levels were lower than in group N, while group C2 showed similar performance to group P1. Groups A2 and P2 had the highest abundance of functional genes related to the biosynthesis of anisoxam and vancomycin-like antibiotics, indicating broad-spectrum inhibition of harmful intestinal bacteria. Furthermore, groups A2, C2, and P2 had the lowest abundance of functional genes related to drug metabolism—specifically, those involved in the biodegradation and metabolism of exogenous compounds and other enzymes. (See [link to relevant documentation]). Figure 8 .

Claims

1. A medicinal and edible composition, characterized in that, The composition is made from 10-30 parts licorice, 20-40 parts buckwheat, and 30-70 parts burdock.

2. The medicinal and edible composition according to claim 1, characterized in that, The composition is made from 10-20 parts licorice, 20-30 parts buckwheat, and 40-70 parts burdock.

3. The medicinal and edible composition according to claim 2, characterized in that, The composition is made from 10-15 parts licorice, 25-30 parts buckwheat, and 55-70 parts burdock.

4. The medicinal and edible composition according to claim 3, characterized in that, The composition is made from 10 parts licorice, 30 parts buckwheat, and 60 parts burdock.

5. A method for preparing the medicinal and edible composition according to any one of claims 1 to 4, characterized in that, According to the mass ratio, weigh out licorice, buckwheat, and burdock seeds, mix them together, dry them to a moisture content of ≤10%, and then pulverize them into powder with a mesh size of ≥300.

6. Feed or feed additive containing any one of the medicinal and edible compositions according to claims 1 to 4.

7. The use of any one of the food-medicine homologous compositions of claims 1 to 4 in the preparation of feed or medicines that improve pig growth performance, stress resistance, liver performance, immunity, and intestinal flora.

8. The application according to claim 7, characterized in that, The medicinal and edible homology composition is added to feed at a rate of 0.1% to 1%.

9. The application according to claim 8, characterized in that, The medicinal and edible homology composition is added to feed at a rate of 0.3%.

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