Plant extract feed additive for regulating intestinal flora of mutton sheep and application of plant extract feed additive

Abstract

The invention relates to the technical field of biology, in particular to a plant extract feed additive for regulating intestinal flora of mutton sheep and application. According to the mutton sheep daily ration, the piper sarmentosum extract is added into the formula of the mutton sheep daily ration, it is proved that the diversity of ileum flora can be improved, the abundance of ileum cellulolytic bacteria can be adjusted, and the adjusting effects include the improvement of the abundance of Proteobacteria gate, the improvement of the abundance of Bacteroidia gate and the improvement of the abundance of Fibrobacteroidia gate and the reduction of the abundance of Fibroidia gate. The intestinal mucosa morphology and the barrier function are effectively improved by adjusting the diversity of the intestinal microorganisms of the mutton sheep and improving the balance of the intestinal microbial flora and the intestinal metabolic map, and the composition has important significance in the field of mutton sheep breeding.

Description

Technical Field

[0001] The invention relates to the biological field, and in particular to a plant extract feed additive for regulating intestinal flora of mutton sheep and its application. Background Art

[0002] As climate change intensifies, many regions are facing increasingly frequent and severe drought events, posing a serious threat to global livestock and poultry production. Environmental stress issues have quickly become a major concern for Hainan's animal husbandry. Environmental stressors, such as heat stress, are particularly harmful to animal husbandry. Heat stress is a series of damaging stress processes on the animal body caused by a negative balance between the net energy flowing from the animal's body to the surrounding environment and the heat energy produced by the animal due to combined changes in environmental factors and animal characteristics. Studies have confirmed that heat stress can have a serious impact on animal growth and production performance. The adverse effects of heat stress on growth performance are mainly attributed to reduced feed intake, decreased digestibility, and reduced energy utilization efficiency. In a high temperature environment, animals eat less and their digestive system functions are impaired, resulting in the ineffective utilization of ingested nutrients, thereby limiting growth and development. Heat stress directly affects meat quality by impairing protein synthesis and increasing unwanted fat in meat. Animals are stressed in a high temperature environment, and the increased body metabolic rate leads to reduced protein synthesis. In addition, heat stress may also promote fat accumulation, resulting in an increase in the fat content in meat, affecting the taste and nutritional value of meat. Animals exposed to heat stress often have an increased pH in their meat due to glycogen depletion, which increases shear stress, leading to a decrease in tenderness and juiciness, and further affecting the color and flavor of the meat.

[0003] Intestinal health is essential to cope with the adverse effects of heat stress on animal husbandry. The gastrointestinal epithelium of ruminants can protect the host from the intestinal environment and control the absorption and metabolism of nutrients. The gastrointestinal epithelial barrier plays a vital role in maintaining the host's energy metabolism and health. One of the key factors for intestinal health is to maintain proper epithelial barrier function in the gastrointestinal tract. The intestinal barrier protects the host from the mixture of microorganisms, toxins and chemicals in the lumen. The epithelial barrier in the intestinal lumen and mucosal immune system contains nearly 70% of the leukocytes and 80% of the antibody-secreting cells in the body, and interacts with the intestinal flora to maintain intestinal homeostasis and intestinal health. An imbalance in the intestinal microbial flora will lead to underdevelopment of the intestinal-related immune system and peripheral organs, and is associated with chronic metabolic disorders, inflammatory bowel disease and autoimmune diseases. Since the ban on antibiotics, low-cost, green, healthy and effective plant extracts that promote intestinal health and improve animal production performance have become an urgent need in the animal husbandry field.

[0004] Hainan's tropical monsoon climate also brings rich tropical germplasm resources, which have broad prospects in the development and utilization of new feed additive products. Piper sarmentosumRobx belongs to the Piperaceae, Piperales, and Piper genus. It is a tropical and subtropical characteristic plant resource with great development potential. It is widely distributed in the southeastern coastal areas of my country, northeastern India, and Southeast Asia, and is commonly used in traditional medicine and food seasoning (Li Jinwei et al., 2017; Aslam et al., 2017). Modern pharmacological studies have shown that Robx extracts have a wide range of biological activities, such as anti-inflammatory and antibacterial, anti-osteoporosis, anti-depression, anti-cancer, hypoglycemic, anthelmintic, antihypertensive, etc. Summary of the invention

[0005] The purpose of the present invention is to provide a plant extract feed additive for regulating the intestinal flora of mutton sheep and its application.

[0006] Plant extracts, as an alternative to antibiotics, have antibacterial and antiviral effects. This alternative can not only effectively reduce the human health risks caused by the abuse of antibiotics, but also help control pathogenic microorganisms in livestock and poultry and improve animal immunity. At the same time, the rich nutrients in plant extracts promote the growth and development of livestock and poultry, and improve product quality and yield. This alternative is more environmentally friendly, reduces the negative impact of antibiotics on the environment, and helps build a sustainable breeding system.

[0007] The invention provides a method for regulating the digestion and absorption capacity of mutton sheep, and uses the Pseudoconopsis konjac extract as a feed additive in the mutton sheep breeding process.

[0008] Furthermore, the Pseudocongolonia extract is a Pseudocongolonia ethanol extract.

[0009] Furthermore, the dosage of the Pseudoconazole extract is 200 mg / kg to 600 mg / kg.

[0010] Furthermore, the dosage of the Pseudoconazole extract is 400 mg / kg.

[0011] Furthermore, the regulation of the digestion and absorption capacity of mutton sheep is to increase the abundance of fiber-degrading bacteria and potential probiotics, and reduce the abundance of conditional pathogens.

[0012] Furthermore, the method for regulating the digestion and absorption capacity of mutton sheep is: increasing Proteobacteria Door, Bacteroidota Door and Fibrobacterota Gate abundance; reduce Firmicutes Door abundance.

[0013] Furthermore, the regulation of the digestion and absorption capacity of mutton sheep is to effectively improve the morphology of the small intestinal mucosa.

[0014] Furthermore, the improvement of small intestinal mucosal morphology includes: improving the morphology of duodenal and ileal mucosal tissues, significantly reducing the depth of ileal crypts, and increasing the ratio of duodenal and ileal villus height to crypt depth.

[0015] Furthermore, the method of regulating the digestion and absorption capacity of mutton sheep also includes: 1) Significantly increased the abundance of genes involved in carbohydrate metabolism, amino acid metabolism, membrane transport, and lipid metabolism at level 2; 2) Significantly improve lipase activity; 3) Improve the intestinal metabolic profile.

[0016] The application of the above method in breeding sheep should also be within the protection scope of the present invention.

[0017] Furthermore, the goats are Hainan black goats, and more specifically, Hainan black goats under heat stress.

[0018] The present invention adds a Pseudoconazole extract (the addition amount is 400 mg / kg) to the daily dietary concentrate of male lambs in the growing and fattening period, regulates the intestinal microbial flora, improves the morphology of the small intestinal mucosa, and promotes intestinal digestion and absorption, thereby maintaining the overall health of the sheep and improving the production efficiency of high-grade mutton.

[0019] The beneficial effects of the present invention are: The present invention provides a plant extract feed additive for regulating the intestinal flora of mutton sheep and its application. For Hainan black goats under heat stress, the method of directly adding Pseudoconazole extract to the feed effectively increases the abundance of fiber-degrading bacteria and potential probiotics, and reduces the abundance of conditional pathogens in Hainan black goats under heat stress, specifically: increasing Proteobacteria Door, Bacteroidota Door and Fibrobacterota Gate abundance; reduce Firmicutes At the same time, it effectively improved the small intestinal mucosal morphology of Hainan black goats under heat stress, specifically: it improved the morphology of duodenal and ileal mucosal tissue, significantly reduced the depth of ileal crypts, and increased the ratio of duodenal and ileal villus height to crypt depth.

[0020] As a common tropical and subtropical plant, the extract of Pseudocong has good sustainability. Through reasonable planting and extraction methods, effective utilization of resources can be achieved, which is conducive to the sustainable development of the environment and ecology. In order to improve intestinal health and promote digestion and absorption, adding natural plant-derived Pseudocong extract to feed instead of commonly used enzyme preparations and compound organic compound preparations has more obvious ecological and social benefits, great promotion value and broad prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 is the average temperature and humidity changes in the sheep house during the experiment.

[0022] Figure 2 The effect of Pseudoconazole extract on intestinal histological morphology of Hainan black goats.

[0023] Figure 3 Effects of Pseudoconazole extract on the microbial flora of the duodenum (A), jejunum (B) and ileum (C) of Hainan black goats.

[0024] Figure 4 Discriminant analysis of LEfSe multi-level species differences in ileal microbial flora from phylum to genus level (LDA>3).

[0025] Figure 5 Effects of Pseudoconazole extract on the ileal microbial flora of Hainan black goats at the phylum level (A), family level (B) and genus level (C).

[0026] Figure 6 Comparison of the abundance of microbiota at the ileum level (A), family level (B) and genus level (C) between the CON and PSE400 groups of Hainan black goats under the influence of Pseudoconazole extract.

[0027] Figure 7 To predict the top 10 differential biological functions at KEGG level 2 between CON and PSE400 groups in the ileal microbiota of Hainan black goats under the influence of Pseudoconazole extract.

[0028] Figure 8 Effect of Pseudoconopsis konjac extract on digestive enzyme activities in ileum of Hainan black goats.

[0029] Fig. 9 OPLS-DA score diagram of CON and PSE400 group samples under the positive ion (A) and negative ion (B) modes of ileal metabolites of Hainan black goats under the influence of Pseudoconazole extract.

[0030] Fig.10 This is a comparison of the differential metabolites in the negative ion mode between the CON and PSE400 groups in the ileal microbiota of Hainan black goats under the influence of Pseudoconazole extract.

[0031] Fig.11 These are the top 20 KEGG pathways enriched in differentially anionic metabolites between CON and PSE400 groups in the ileum of Hainan black goats under the influence of Pseudoconazole extract.

[0032] Fig.12 This is a MetOrigin-based evaluation of the differential metabolites in the fructose and mannose metabolic pathways between CON and PSE400 groups in Hainan black goats’ ileum. BIO-Sankey network of metabolic reactions R01332 (A), R01326 (C), and R02630 (E) in fructose and mannose metabolism. STA-Sankey network of metabolic reactions R01332 (B), R01326 (D), and R02630 (F) in fructose and mannose metabolism. Asterisks (*) indicate statistically significant correlations with metabolites.

[0033] Fig.13 This is a MetOrigin-based evaluation of differential metabolites in the linoleic acid metabolic pathway between CON and PSE400 groups in Hainan black goats’ ileum. BIO-Sankey network of metabolic reactions R07063 (A) and R07064 (C) in linoleic acid metabolism. STA-Sankey network of metabolic reactions R07063 (B) and R07064 (D) in fructose and mannose metabolism. Asterisks (*) indicate statistically significant correlations with metabolites.

[0034] Fig.14 The experimental flow chart is shown in Figure 2. DETAILED DESCRIPTION

[0035] The present invention is further described in detail below in conjunction with specific embodiments, and the examples provided are only for illustrating the present invention, rather than for limiting the scope of the present invention. The examples provided below can be used as a guide for further improvements by those of ordinary skill in the art, and do not constitute a limitation of the present invention in any way.

[0036] The experimental methods in the following examples, unless otherwise specified, are conventional methods, carried out according to the techniques or conditions described in the literature in this field or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial sources. The Hainan black goats in the following examples were purchased from the Chinese Academy of Tropical Agricultural Sciences.

[0037] Example 1: Grouping design of experimental animals Thirty-six healthy weaned purebred Hainan black goats (9.48 ±0.25 kg) of the same strain, three months of age, similar weight, were selected and divided into four groups for feeding. A completely randomized design was adopted, and the 36 goats were randomly divided into 4 groups: control group, 200 mg / kg Pseudocones extract added group (PSE200), 400 mg / kg Pseudocones extract added group (PSE400), and 600 mg / kg Pseudocones extract added group (PSE600). The experimental time was from July 2022 to October 2022. From weaning to the end of fattening, the experiment lasted a total of 105 days, including a pre-trial period of 15 days and a regular trial period of 90 days. The ambient temperature and humidity were recorded daily, and the THI index was calculated. THI=0.8×ambient temperature+(relative humidity / 100)×(ambient temperature-14.4)+46.4. Based on the following Fig.14As shown in the experimental flow chart, the animal feeding experiment was conducted at the Hainan Black Goat Breeding Base in Danzhou City, Hainan Province. The experiment was carried out from July to October. During the experiment, Hainan Province was in summer, with high temperature and high humidity. THI was mostly between 79 and 89, indicating that the experimental animals were in a state of high heat stress. After the experiment, the animals were slaughtered and the small intestine and small intestinal contents were taken for analysis.

[0038] Among them, the addition of pseudocones extract and daily feeding: before the start of the experiment, the feeding pens were disinfected and the test sheep were uniformly dewormed and immunized. The test time is from July 2022 to October 2022. The test lasted for a total of 105 days, including a pre-test period of 15 days and a main test period of 90 days. Feed once at 8:30 and 15:00 every day, and each test sheep is fed with sufficient basic diet and free drinking water. The amount of feed in the pre-test period is based on a slight surplus the next day to determine the amount of feed intake of the sheep. Clean the sheep house on time every week, clean up the feces and disinfect the environment.

[0039] Example 2: Preparation of experimental basal diet and analysis of nutritional levels 1. Preparation of Pseudoconazole Extract Referring to the method described in the literature "Effect of Pseudocongolonia Extract on In Vitro Rumen Fermentation Parameters and Dry Matter Digestibility of Hainan Black Goats [J]. Chinese Journal of Animal Husbandry, 2024, 60(05): 233-238. DOI: 10.19556 / j.0258-7033.20230723-01.", the cold soaking method was used to extract Pseudocongolonia powder with 95% ethanol and rotary evaporation to obtain Pseudocongolonia crude ethanol extract, of which polyphenols and flavonoids accounted for 27.5%; after freeze-drying into powder, it was evenly mixed with zeolite powder in a volume ratio of 1:1 and stored at 4°C for later use.

[0040] 2. Feed configuration The basic diet was formulated according to the nutritional requirements of mutton sheep (NY / T 816-2021). In the experimental basic diet, the concentrated feed used corn-soybean meal type diet, and the roughage used king grass. The dry matter, energy, crude protein, neutral detergent fiber, acid detergent fiber, calcium, phosphorus, etc. in the feed were determined according to the method of Zhang Liying (2003). Its composition and nutritional level are shown in Table 1 below.

[0041] Table 1 Basic diet composition and nutritional level (dry matter basis)

[0042] 1) The premix provides 15000 IU of vitamin A, 5000 IU of vitamin D, 50 mg of vitamin E, 9 mg of iron, 12.5 mg of copper, 100 mg of zinc, 130 mg of manganese, 0.3 mg of selenium, 1.5 mg of iodine and 0.5 mg of cobalt per kilogram of feed.

[0043] 2) Except for digestible energy, all nutritional levels are measured values.

[0044] Example 3: Effect of dietary supplementation with Pseudo-conch extract on intestinal histological morphology of mutton sheep Small intestinal segments (duodenum, jejunum, and ileum) were collected, gently rinsed with phosphate buffer solution, and fixed with 4% paraformaldehyde. They were then dehydrated, paraffin-embedded, sectioned, and stained with hematoxylin and eosin (HE). Intestinal images were obtained under a 40x light microscope. Six typical fields of view with intact and straight villi were selected on the tissue sections using NDP.view2 software to measure the height of the villi and the depth of the crypts connected to them. The results of intestinal histological measurements were statistically tested using IBM SPSS Statistics 25. One-way ANOVA was used to test the differences between treatment groups, and Tukey's multiple comparison method was used to test significant differences.

[0045] As shown in Table 2, Figure 2 As shown in the results, the addition of Pseudo-cone extract to the diet effectively improved the morphology of the duodenal and ileal mucosal tissues, significantly reduced the depth of the ileal crypts, and increased the ratio of the villus height to the crypt depth in the duodenum and ileum ( P <0.05).

[0046] Table 2 Effects of Pseudoconazole extract on intestinal histological parameters of Hainan black goats

[0047] Note: The same letters in the same row indicate no significant difference, while different letters indicate significant difference. The same below.

[0048] Example 4: Effect of adding Pseudoconazole extract to the diet on intestinal microorganisms of mutton sheep HiPure Stool DNA Kits (Magen Biotechnology Co., Ltd., Guangzhou, China) were used to extract total microbial DNA. The purity and integrity of nucleic acids were tested using NanoDrop 2000 (Thermo Fisher Science, Wilmington, DE, USA) and DYY-6C agarose gel electrophoresis (Liuyi Biotechnology Co., Ltd., Beijing, China). Then, PCR (V3-V4 region) amplification, purification, and quantification were performed according to the procedures of Zhan et al. (2019). The second round of amplification products were purified using AMPure XP Beads, quantified using ABI StepOnePlus Real-Time PCR System (Life Technologie, USA), and sequenced using Novasek 6000 PE250 mode pooling. UPARSE (version 9.2.64), QIIME (version 1.9.1), and Muscle (version 3.8.31) were used to perform bioinformatics analysis (OUT, community composition, α diversity, β diversity analysis, and functional prediction) of the raw data. All sequencing and bioinformatics analyses were performed using the Omicsmart online platform (http: / / www.omicsmart.com).

[0049] Table 3 Figure 3 The addition of 400 or 600 mg / kg of Pseudocong extract to the diet changed the microbial flora of each intestinal segment, and 400 mg / kg of Pseudocong extract significantly increased the ileal microbial diversity ( P <0.05).

[0050] Table 3. Effects of Pseudoconazole extract on the Alpha diversity of intestinal microorganisms in Hainan black goats

[0051] Figure 4 At the phylum level, the dominant phylum in the ileum of Hainan black goat is Firmicutes Door and Proteobacteria At the genus level, the main genera are Acinetobacter Genus, Solibacillus Genus and Lysinibacillus Belong.

[0052] Figure 5The differential biomarkers closely related to the improvement of intestinal barrier function by Pseudoconch extract were further determined by LEfSe analysis of the evolutionary branch diagram. The results showed that the Con group, PSE200 group, PSE400 group and PSE600 group contained 34, 8, 26 and 9 statistically significant biomarkers (LDA>3), respectively. The biomarkers of the CON group were Actinobacteriota Door, Chloroflexi Door and Cyanobacteria The biomarkers of the PSE400 group were Bacteroidota Door and Fibrobacterota Door.

[0053] Figure 6 The abundance of fiber-degrading bacteria in the ileum PSE400 group was significantly changed compared with the CON group ( P <0.05), specifically the PSE400 group Proteobacteria Door, Bacteroidota Door and Fibrobacterota The abundance of the phylum increased significantly. Firmicutes The abundance of the phylum was significantly reduced. Bacteroidota Door and Firmicutes All phyla play an important role in the degradation and utilization of intestinal fiber and carbohydrates. Bacteroidota The phylum has a variety of enzymes that can utilize glycans and glycosides of polysaccharides, and their dynamic changes may reflect adaptation to complex carbohydrates. Acinetobacter Genus, Bacteroidales_bacterum_bact_22 Genus, Rikenellaceae_RC9_gut_group Genus, Prevotella Genus, Prevotellaceae_UCG-001 Genus, Prevotellaceae_UCG-003 Genus and UCG-005 The relative abundance of the genera increased, confirming the increased capacity for fiber and carbohydrate degradation and utilization in the ileal PSE400 group.

[0054] like Figure 7 The results of KEGG functional analysis showed that the addition of 400 mg / kg of Pseudoconch extract to the ileum significantly increased the gene abundance of level 2 carbohydrate metabolism, amino acid metabolism, membrane transport, and lipid metabolism. P <0.05). Membrane transport is essential for bacteria to absorb and transport monosaccharide substrates for the production of SCFA. Bacteria can also produce propionic acid and butyric acid as fermentation byproducts by taking up acetic acid. This indicates that the addition of Pseudoconazole extract improves the ability of the flora to metabolize carbohydrates and increase SCFA production, thereby compensating for the ability of the intestinal flora to utilize SCFA.

[0055] Example 5: Effect of dietary supplementation with Pseudoconazole extract on ileal metabolic profile of mutton sheep The collected ileal contents were homogenized in saline and then centrifuged at 2000 × g for 10 min at 4 °C. The supernatant was collected and immediately frozen at −80 °C for later analysis. α-Amylase, cellulase, chymotrypsin, trypsin, and lipase activities were determined according to the manufacturer's instructions (Nanjing Jiancheng Bioengineering Institute, Nanjing, China).

[0056] After the ileum contents were slowly thawed at 4°C, 100 μL of each aliquot was taken and added to 400 μL of pre-cooled methanol / acetonitrile solution (1:1, v / v), vortexed, allowed to stand at -20°C for 30 min, centrifuged at 14,000 g for 20 min at 4°C, the supernatant was taken, vacuum dried, and for mass spectrometry analysis, 100 μL of acetonitrile aqueous solution (acetonitrile: water = 1:1, v / v) was added for reconstitution, vortexed, and centrifuged at 14,000 g for 15 min at 4°C. The supernatant was taken for sampling and analysis.

[0057] Agilent 1290 Infinity LC ultra high performance liquid chromatography (UHPLC) system with HILIC column was used for separation; column temperature was 25℃; flow rate was 0.5 mL / min; injection volume was 2 μL; mobile phase composition A: water + 25 mM ammonium acetate + 25 mM ammonia, B: acetonitrile; gradient elution program was as follows: 0--- 0.5 min, 95% B; 0.5---7 min, B changed linearly from 95% to 65%; 7---8 min, B changed linearly from 65% to 40%; 8--- 9 min, B was maintained at 40%; 9---9.1 min, B changed linearly from 40% to 95%; 9.1--- 12 min, B was maintained at 95%; the sample was placed in an autosampler at 4℃ during the entire analysis process.

[0058] An AB Triple TOF 6600 mass spectrometer was used to collect the primary and secondary spectra of the samples.

[0059] The ESI source conditions after HILIC chromatographic separation are as follows: Ion Source Gas1 (Gas1): 60, Ion SourceGas2 (Gas2): 60, Curtain gas (CUR): 30, source temperature: 600℃, IonSapary VoltageFloating (ISVF) ±5500 V (positive and negative modes); TOF MS scan m / z range: 60-1000 Da, product ion scan m / z range: 25-1000 Da, TOF MS scan accumulation time 0.20 s / spectra, product ion scan accumulation time 0.05 s / spectra; The secondary mass spectrum was obtained using information dependent acquisition (IDA) in high sensitivity mode, Declustering potential (DP): ±60 V (positive and negative modes), Collision Energy: 35±15 eV, IDA settings are as follows: Exclude isotopes within 4 Da, Candidate ions to monitor per cycle: 10. Bioinformatics analysis was performed using the Omicsmart online platform (http: / / www.omicsmart.com). The origin of ileal metabolites was assessed based on the MetOrigin database (2024-08 version, Capra hircus, https: / / metorigin.metbioinformatics.cn).

[0060] like Figure 8 As shown in Figure 2, due to the observed changes in the composition and functional characteristics of the ileal microbiota, we measured the digestive enzyme activities in the ileal contents for verification. The results showed that the lipase activity in the PSE400 group was significantly increased compared with the CON group ( P< 0.05); while the cellulase activity was not statistically significant, but the results in the PSE400 and PSE600 groups were close to the significant level (0.1> P >0.05) Figure 9-11Further metabolomics results showed that the ileal metabolites of the CON and PSE400 groups were clearly separated. Given that our main focus was on carbohydrates and lipids, which were more effectively ionized and detected in negative ion mode, we chose to focus on the negative ion mode. Through the VIP bubble plot, we identified the key metabolites that differed between the conditions observed between the CON and PSE400 groups. In negative ion mode, the key metabolites were mainly fatty acyl groups (azelaic acid, prostaglandin I2, linoleic acid, linolenic acid, and heptadecanoic acid). Compared with the CON group, the PSE400 group showed significant changes mainly in the biosynthesis of unsaturated fatty acids, fructose and mannose metabolism, linoleic acid metabolism, antifolate resistance, vitamin digestion and absorption, biosynthesis of plant secondary metabolites, and benzoate degradation. P<0.05). Metabolomic analysis showed that PSE increased the concentrations of eicosenoic acid, linoleic acid, and linolenic acid, which are associated with the synthesis of unsaturated fatty acids, and 13-kode and linoleic acid, which are associated with linoleic acid metabolism. These unsaturated fatty acids play a role in reducing inflammation, supporting cell membranes, and maintaining overall metabolic balance. Many studies have highlighted the potential health benefits of increasing n-3 polyunsaturated fatty acid intake, including improved neurological and cardiovascular health and reduced inflammation. 13-kode exhibits potent anti-inflammatory and antioxidant activities by inhibiting nitric oxide production, suppressing the expression of proinflammatory cytokines, and reducing oxidative stress by regulating key signaling pathways. PSE affects the synthesis and metabolic pathways of unsaturated fatty acids, which may help improve overall metabolic function and prevent oxidative stress. In ruminants, the ileum of the small intestine is the main site of fatty acid absorption, and the fatty acid profile may change with the increase of unsaturated fatty acids. This change may affect the nutritional value of meat, and higher levels of unsaturated fatty acids may improve the health attributes of meat, such as reducing the risk of cardiovascular disease. In addition, PSE increased the concentration of D-mannose, which is associated with fructose and mannose metabolism, while sorbitol 6-phosphate was significantly reduced. D-mannose is a monosaccharide with significant biological functions, which can effectively prevent bacterial adhesion and colonization in animal models, serve as an alternative to antibiotics in animal feed, and promote animal health by inhibiting the growth of intestinal pathogens.

[0061] Figure 12-13 Metorigin-based combined statistical and biological correlation analysis confirmed that Proteobacteria Door, Bacteroidota Door and Fibrobacterota The increase of doors, Firmicutes The reduction of gate was the main reason for the difference in metabolic profiles between the CON and PSE400 groups.

[0062] It can be seen that adding the extract of Pseudocongol in the feed can improve the digestion and absorption capacity and the intestinal metabolic profile by regulating the structure of intestinal microbial flora and improving the morphology of small intestinal mucosa. This shows that the feed containing the extract of Pseudocongol provided by the present invention has certain potential value in improving the intestinal health of mutton sheep.

[0063] The present invention has been described in detail above. It will be apparent to those skilled in the art that the present invention may be implemented in a wide range under equivalent parameters, concentrations and conditions without departing from the spirit and scope of the present invention and without the need for unnecessary experimentation. Although the present invention provides specific embodiments, it should be understood that further improvements may be made to the present invention. In short, according to the principles of the present invention, this application intends to include any changes, uses or improvements to the present invention, including changes made by conventional techniques known in the art that depart from the scope disclosed in this application. Applications of some of the basic features may be made within the scope of the following appended claims.

Claims

1. A method for regulating the digestion and absorption capacity of mutton sheep, characterized in that: Use of Pseudoconazole extract as a feed additive in the feeding process of mutton sheep.

2. The method for regulating the digestion and absorption capacity of mutton sheep according to claim 1, characterized in that: The Pseudocongia konjac extract is Pseudocongia konjac ethanol extract.

3. The method for regulating the digestion and absorption capacity of mutton sheep according to claim 2, characterized in that: The dosage of the Pseudoconazole extract is 200 mg / kg to 600 mg / kg.

4. The method for regulating the digestion and absorption capacity of mutton sheep according to claim 3, characterized in that: The dosage of the Pseudocongia konjac extract is 400 mg / kg.

5. The method for regulating the digestion and absorption capacity of mutton sheep according to claim 1, characterized in that: The method for regulating the digestion and absorption capacity of mutton sheep is to regulate the abundance of fiber-degrading bacteria.

6. The method for regulating the digestion and absorption capacity of mutton sheep according to claim 5, characterized in that: The method for regulating the digestion and absorption capacity of mutton sheep is: increasing Proteobacteria Door, Bacteroidota Door and Fibrobacterota Gate abundance; reduce Firmicutes Door abundance.

7. The method for regulating the digestion and absorption capacity of mutton sheep according to claim 1, characterized in that: The regulation of the digestion and absorption capacity of mutton sheep is to effectively improve the morphology of the small intestinal mucosa.

8. The method for regulating the digestion and absorption capacity of mutton sheep according to claim 7, characterized in that: The improvement of small intestinal mucosal morphology includes: improving the morphology of duodenal and ileal mucosal tissues, significantly reducing the depth of ileal crypts, and increasing the ratio of duodenal and ileal villus height to crypt depth.

9. The method for regulating the digestion and absorption capacity of mutton sheep according to claim 1, characterized in that: The regulation of the digestion and absorption capacity of mutton sheep also includes: 1) Significantly increased the abundance of genes involved in carbohydrate metabolism, amino acid metabolism, membrane transport, and lipid metabolism at level 2; 2) Significantly improve lipase activity; 3) Improve the intestinal metabolic profile.

10. Use of the method according to any one of claims 1 to 9 in breeding sheep.

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