A microecological preparation, a composite preparation thereof and application thereof

Abstract

The present application belongs to the field of microbial technology, and particularly relates to a micro-ecological preparation, a composite preparation thereof and application. The present application discloses a micro-ecological preparation and a composite preparation thereof. The composite preparation comprises the micro-ecological preparation and a composite enzyme preparation. The micro-ecological composite preparation comprises Lactobacillus plantarum CGMCC NO.12849, Bacillus subtilis CGMCC NO.25499 and Saccharomyces cerevisiae CGMCC NO.21678. The composite enzyme preparation comprises cellulase, xylanase and pectinase respectively. The endotoxin-degrading bacterial enzyme composite preparation realizes that the endotoxin degradation rate of feed reaches more than 95%, and solves the problems of animal health and production loss caused by feed endotoxin pollution.

Description

Technical Field

[0002] This invention belongs to the field of feed microecological technology, specifically relating to a microecological preparation and its compound preparation and application. Background Technology

[0004] Bacterial endotoxins, also known as lipopolysaccharides (LPS), are widely found on the outer membrane of the cell walls of Gram-negative bacteria such as Escherichia coli, Salmonella, Brucella, Salmonella typhi, and Proteus. The LPS molecule consists of three parts: lipid A, core polysaccharide, and O antigen. Lipid A is the hydrophobic anchoring group of the endotoxin and the core structural basis for its biotoxic activity. During conventional feed production, environmental stimuli and processing techniques can introduce a certain amount of bacterial endotoxins, leading to reduced feed quality and threatening animal health. Piglets, in particular, are susceptible to multiple stressors after weaning, resulting in weakened immunity and increased risk of endotoxin poisoning. While existing feed processing techniques (such as high-temperature pelleting and extrusion) can effectively kill live bacteria, they cannot destroy the extremely stable chemical structure of endotoxins (especially the heat-resistant lipid A). Conversely, during sterilization, bacterial death, autolysis, or cell wall rupture can cause large amounts of endotoxins originally bound to the cell membrane to be released into the feed matrix. This "dead bacteria release" effect means that although high-temperature processed feed is sterile, it often contains high concentrations of free endotoxins. While the intestines of healthy animals have a barrier function, long-term exposure to high concentrations of feed-derived endotoxins disrupts the tight junctions of intestinal mucosal epithelial cells, significantly increasing intestinal permeability. Subsequently, endotoxins can breach the intestinal barrier, translocate into the bloodstream and lymphatic system, activate the Toll-like receptor 4 (TLR4) signaling pathway on host immune cells, triggering a systemic inflammatory cascade response that threatens animal production and health.

[0005] Currently, the main methods for treating endotoxins in feed on the market include (1) adding chemical adsorbents such as bentonite and activated carbon, which have limited adsorption capacity and selectivity for endotoxins and are easily combined with other nutrients in the feed, affecting feed utilization; (2) adding enzyme preparations, such as lipopolysaccharide hydrolase, but the activity of single enzyme preparations in complex feed systems is easily inhibited by factors such as pH, temperature and metal ions; (3) applying probiotics, mainly lactic acid bacteria or yeast, which reduce the biological activity of endotoxins through competitive colonization and release of metabolites, but the efficacy of a single strain is often limited to specific environmental conditions. Therefore, it is crucial to construct a multifunctional, synergistic compound microbial preparation that can simultaneously exert the synergistic effect of different strains and enzymes, efficiently and stably degrade endotoxins in feed, and at the same time have certain probiotic effects such as disease resistance, disease prevention and promotion of animal growth. Summary of the Invention

[0007] To address the aforementioned technical problems, the present invention aims to provide a microecological preparation, which is made from Lactobacillus plantarum CGMCC NO. 12849, Bacillus subtilis CGMCC NO. 25499, and Saccharomyces cerevisiae CGMCC NO. 21678.

[0008] Furthermore, the microecological preparation is made from Lactobacillus plantarum CGMCC NO. 12849 bacterial powder, Bacillus subtilis CGMCC NO. 25499 bacterial powder, and Saccharomyces cerevisiae CGMCC NO. 21678 bacterial powder, wherein the viable count of the bacterial powder is 1×10⁻⁶. 9 ~1×10 11 CFU / g.

[0009] Preferably, the viable count of the *Lactobacillus plantarum* CGMCC NO. 12849 bacterial powder is 1 × 10⁻⁶. 10 The viable count of the Bacillus subtilis CGMCC NO.25499 bacterial powder was 6 × 10⁻⁶ CFU / g. 9 The viable count of the yeast CGMCC NO.21678 powder was 3 × 10⁻⁶ CFU / g. 9 CFU / g.

[0010] Furthermore, the microecological preparation is made from Lactobacillus plantarum CGMCC NO.12849 bacterial powder, Bacillus subtilis CGMCC NO.25499 bacterial powder and Saccharomyces cerevisiae CGMCC NO.21678 bacterial powder in a mass ratio of (0.5~2):(1~5):(0.5~2), preferably 1:5:1.

[0011] Furthermore, the compound preparation also includes a compound enzyme preparation, wherein the mass ratio of the microecological preparation to the compound enzyme preparation is 13:7.

[0012] Furthermore, the compound enzyme preparation includes cellulase, xylanase, and pectinase, wherein the mass ratio of cellulase, xylanase, and pectinase is 5:1:1.

[0013] The present invention also provides a fermented feed, which is made by fermentation of the aforementioned compound preparation.

[0014] The present invention also provides a method for preparing the fermented feed, wherein 3% to 7% of a compound preparation and 10% molasses are added to the feed raw materials, the moisture content of the fermented feed is controlled to be 50% to 60%, the pH is 5.5 to 6.0, and the feed is cultured at 30°C for 3 to 4 days.

[0015] Furthermore, the feed ingredients include Escherichia coli single-cell protein and wheat bran, wherein the mass ratio of Escherichia coli single-cell protein to wheat bran is 4:1.

[0016] The present invention also provides the application of the microecological preparation or the compound preparation in the degradation of endotoxins in feed, the application including: adding the preparation to feed by fermentation or non-fermentation.

[0017] Preferably, the microecological preparation or the compound preparation is used to ferment the feed to reduce the endotoxin content in the feed.

[0018] The present invention also provides the application of the microecological preparation or the compound preparation in the preparation of feed that improves the intestinal health of animals.

[0019] Furthermore, the improvement of gut health includes at least one of the following: reducing serum endotoxins, inhibiting serum inflammatory factors, and improving colon length.

[0020] Furthermore, the application can also achieve at least one of the following effects: improved growth performance, reduced serum C-reactive protein and lactate dehydrogenase levels, improved spleen coefficient, and improved liver coefficient.

[0021] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0022] This invention achieves a feed endotoxin degradation rate of over 95% through a specific combination of Lactobacillus plantarum CGMCC NO.12849, Bacillus subtilis CGMCC NO.25499, and Saccharomyces cerevisiae CGMCC NO.21678, and synergistic effects with a complex enzyme. Simultaneously, it reduces serum endotoxins, inhibits serum inflammatory factors, improves colon length, and significantly improves animal growth performance. Furthermore, the process is widely adaptable and systematically solves the problem of animal health and production losses caused by feed endotoxin contamination. Attached Figure Description

[0024] Figure 1 Comparison of colon length in healthy mice before and after feeding them with a diet treated with a bacterial enzyme compound preparation.

[0025] Figure 2 The change in body weight of mice with enteritis in different treatment groups over 28 days.

[0026] Figure 3 Changes in blood C-reactive protein levels in mice with enteritis before and after feeding them with a bacterial enzyme compound preparation.

[0027] Figure 4 Changes in blood lactate dehydrogenase levels in mice with enteritis before and after feeding them with a bacterial enzyme compound preparation.

[0028] Figure 5Comparison of colon length in mice with enteritis before and after feeding them with a bacterial enzyme compound preparation. Detailed Implementation

[0030] 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 conventional products.

[0031] Example 1: Preparation of Fermented Feed with Added Degradable Endotoxin Bacterial Enzyme Compound

[0032] 1. Culture medium preparation:

[0033] MRS liquid culture medium consists of the following components and concentrations: yeast extract 5 g / L, glucose 20 g / L, dipotassium hydrogen phosphate 2 g / L, diammonium citrate 2 g / L, sodium acetate 3 g / L, magnesium sulfate 0.2 g / L, manganese sulfate 0.05 g / L, and Tween 80 1 mL / L.

[0034] LB liquid culture medium consists of the following components and concentrations: yeast extract 5 g / L, peptone 10 g / L, and sodium chloride 10 g / L.

[0035] YEPD liquid culture medium, with the following composition and content: peptone 20g / L, yeast extract 10g / L, glucose 20g / L.

[0036] 2. Primary activation:

[0037] Single colonies of *Lactobacillus plantarum* CGMCC NO.12849, *Bacillus subtilis* CGMCC NO.25499, and *Saccharomyces cerevisiae* CGMCC NO.21678 were inoculated into MRS liquid medium, LB liquid medium, and YEPD liquid medium, respectively, and cultured at 37℃, 32℃, and 30℃ for 18 h to obtain primary seed culture.

[0038] 3. Fermentation broth preparation:

[0039] Primary seed cultures of Lactobacillus plantarum CGMCC NO.12849, Bacillus subtilis CGMCC NO.25499, and Saccharomyces cerevisiae CGMCC NO.21678 were inoculated into MRS liquid medium, LB liquid medium, and YEPD liquid medium, respectively, and cultured at 37℃, 32℃, and 30℃ for 48 h to obtain fermentation broth.

[0040] 4. Preparation of microbial agents:

[0041] Lactobacillus plantarum CGMCC NO.12849 fermentation broth was freeze-dried to prepare Lactobacillus plantarum inoculum, with a viable count of 1×10⁻⁶ cells. 10 CFU / g; Bacillus subtilis inoculum was prepared by spray drying the fermentation broth of Bacillus subtilis CGMCC NO.25499, with a viable count of 6 × 10⁻⁶. 9 CFU / g; Saccharomyces cerevisiae CGMCC NO.21678 fermentation broth was freeze-dried and then treated to prepare a Saccharomyces cerevisiae inoculum, with a viable count of 3 × 10⁻⁶ CFU / g; Saccharomyces cerevisiae CGMCC NO.21678 fermentation broth was freeze-dried and then treated to obtain a Saccharomyces 9 CFU / g.

[0042] Among them, *Lactobacillus plantarum* CGMCC NO.12849 is a strain owned by our laboratory. This strain is named *Lactobacillus plantarum* DBNZW02 and was deposited on August 15, 2016, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, with accession number CGMCC NO.12849. For details, please refer to patent CN108570421B.

[0043] Bacillus subtilis CGMCC NO.25499 is a strain owned by our laboratory. This strain is named Bacillus subtilis BS-ZPW and was deposited on August 8, 2022, at the China General Microbiological Culture Collection Center, located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, with accession number CGMCC NO.25499. For details, please refer to patent CN 115181707 A.

[0044] Saccharomyces cerevisiae CGMCC NO.21678 is a strain owned by our laboratory and was deposited on January 19, 2021, at the China General Microbiological Culture Collection Center, located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, with the accession number CGMCC NO.21678. See patent CN 114891650 B for details.

[0045] 5. Endotoxin degradation protocol and effect evaluation:

[0046] Lactobacillus plantarum CGMCC NO.12849, Bacillus subtilis CGMCC NO.25499, and Saccharomyces cerevisiae CGMCC NO.21678 were individually or in different mass ratios to obtain microbial compound preparations. These microbial compound preparations were then thoroughly mixed with a compound enzyme preparation (cellulase:xylanase:pectinase = 5:1:1) at a mass ratio of 13:7 to obtain a microbial-enzyme compound preparation. This microbial-enzyme compound preparation was added to feed ingredients (Escherichia coli single-cell protein: wheat bran = 4:1), with the microbial-enzyme compound preparation accounting for 5% of the dry weight of the feed ingredients, including 10% molasses. The fermented feed had a moisture content of 50%–60% and a pH of 5.5–6.0. ​​The feed was cultured at 30℃ for 3–4 days to obtain fermented feed. The fermented feed was evaluated for endotoxins using the Limulus amebocyte lysate (LAL) reagent-dynamic turbidimetric assay (kit).

[0047] The endotoxin degradation grouping scheme and proportions are as follows:

[0048] (1) Lactobacillus plantarum CGMCC NO.12849 fermentation inoculum;

[0049] (2) Bacillus subtilis CGMCC NO.25499 fermentation inoculum;

[0050] (3) Brewing yeast CGMCC NO.21678 fermentation inoculum;

[0051] (4) Lactobacillus plantarum CGMCC NO.12849: Bacillus subtilis CGMCC NO.25499 = 1:1;

[0052] (5) Lactobacillus plantarum CGMCC NO.12849: Saccharomyces cerevisiae CGMCC NO.21678 = 1:1;

[0053] (6) Bacillus subtilis CGMCC NO.25499: Saccharomyces cerevisiae CGMCC NO.21678 = 1:1;

[0054] (7) Lactobacillus plantarum CGMCC NO.12849: Bacillus subtilis CGMCC NO.25499: Saccharomyces cerevisiae CGMCC NO.21678 = 1:1:1;

[0055] (8) Lactobacillus plantarum CGMCC NO.12849: Bacillus subtilis CGMCC NO.25499: Saccharomyces cerevisiae CGMCC NO.21678 = 1:3:1;

[0056] (9) Lactobacillus plantarum CGMCC NO.12849: Bacillus subtilis CGMCC NO.25499: Saccharomyces cerevisiae CGMCC NO.21678 = 1:5:1;

[0057] The endotoxin test results of the above treatments are shown in Table 1. The results show that scheme 9, namely Lactobacillus plantarum CGMCC NO.12849: Bacillus subtilis CGMCC NO.25499: Saccharomyces cerevisiae CGMCC NO.21678 = 1:5:1, has the best degradation effect.

[0058] Table 1. Results of endotoxin degradation rates in different groups

[0059]

[0060] Example 2: Application of endotoxin-degrading bacterial enzyme complex in healthy mice

[0061] Forty-five female KM mice aged 4-6 weeks were used as experimental animals. The mice were divided into three treatment groups: a control group, a model group, and an experimental group, with five mice per cage and three cages per group. Among them:

[0062] The control group was fed a standard rodent diet, which was a commercially available product with a nutritional composition of 60-70% carbohydrates, 15-20% protein, and 4-7% fat.

[0063] The model group was fed normal rat food plus the mixed feed ingredients from Example 1 that were not treated with the endotoxin-degrading bacterial enzyme compound preparation. The mass ratio of normal rat food to the mixed feed ingredients from Example 1 that were not treated with the endotoxin-degrading bacterial enzyme compound preparation was 4:1.

[0064] The experimental group was fed normal rat food plus the mixed feed ingredients treated with the endotoxin-degrading bacterial enzyme compound preparation of Scheme 8 in Example 1. The mass ratio of normal rat food to the mixed feed ingredients treated with the endotoxin-degrading bacterial enzyme compound preparation of Scheme 8 in Example 1 was 4:1.

[0065] Mice were pre-fed for 3 days, with free access to food and water, for a total of 4 weeks. The mice's condition and mortality were observed weekly, and their weight and food intake were recorded. After 4 weeks of feeding, blood was collected from the eyeballs. The collected whole blood was allowed to stand at 4°C for 12 hours, centrifuged at 4000 rpm for 20 minutes, and the supernatant was collected. The serum endotoxin content was determined using a dynamic turbidimetric method. Serum levels of inflammatory factors TNF-α and IL-1β were measured using a kit. At the end of the experiment, the mice were anesthetized, euthanized by cervical dislocation, and the abdominal cavity was opened. All organs were removed and weighed, and the entire colon was harvested, its length recorded, and photographed.

[0066] 1. Behavioral performance and mortality rate of mice

[0067] The mice in the control group exhibited normal activity, thick, smooth, and glossy fur, were responsive, had normal fecal morphology, and were in good condition; no mice died during the 28-day experiment. The mice in the model group exhibited normal activity, smooth fur, were responsive, had normal fecal morphology, and increased anxiety-like behavior; no mice died during the 28-day experiment. The mice in the experimental group exhibited normal activity, normal fecal morphology, smooth fur, were responsive, and were in good condition; no mice died during the 28-day experiment.

[0068] 2. Growth performance

[0069] Table 2 Growth performance of mice in different treatment groups

[0070]

[0071] Note: Different letters indicate significant differences (p < 0.05) in the same column.

[0072] The growth performance of mice after 28 days of feeding is shown in Table 2. Compared with the model group, the experimental group showed significant advantages in weight gain and feed conversion ratio.

[0073] 3. Endotoxin levels and serum inflammatory factors

[0074] Table 3. Plasma endotoxin levels and serum inflammatory mediator levels in mice under different treatment groups.

[0075]

[0076] Note: Different letters indicate significant differences (p < 0.05) in the same column.

[0077] After 28 days of feeding, the levels of endotoxins in the blood and serum inflammatory mediators in mice are shown in Table 3. Compared with the model group, the experimental group showed significantly lower levels of endotoxins, serum inflammatory factors TNF-α and IL-1β.

[0078] 4. Organ coefficient

[0079] Table 4 Comparison of organ coefficients in mice from different treatment groups

[0080] Table 4 shows the comparison of organ coefficients in mice from different treatment groups after 28 days of the experiment. Compared with the model group, the spleen coefficient, liver coefficient, and kidney coefficient were all significantly reduced in the experimental group, while the colon length was not significantly different from that in the control group (see Table 4). Figure 1 ).

[0081] Example 3: Application of endotoxin-degrading bacterial enzyme complex in mice with enteritis

[0082] The experimental animals were 4-6 week old female wild-type C57BL / 6 mice. Mice were randomly divided into groups of 5 mice per cage and 3 cages per group. The mice were further divided into a control group, a model group, a pre-degradation experimental group, and a post-degradation experimental group. All groups were pre-fed normal feed for 3 days. The treatments for each group were as follows:

[0083] The control group always drank deionized water and was fed normal rodent food;

[0084] The model group drank deionized water containing 2.5% DSS and was fed normal rodent food;

[0085] Before treatment, the experimental group drank deionized water containing 2.5% DSS and was fed normal rat food plus the mixed feed ingredients from Example 1 that were not treated with the endotoxin-degrading bacteria enzyme compound preparation. The mass ratio of normal rat food to the mixed feed ingredients from Example 1 that were not treated with the endotoxin-degrading bacteria enzyme compound preparation was 4:1.

[0086] After degradation, the experimental group drank deionized water containing 2.5% DSS and was fed normal rat food plus a mixed feed ingredient treated with the endotoxin-degrading bacteria and enzyme compound preparation in Scheme 8 of Example 1. The mass ratio of normal rat food to the mixed feed ingredient treated with the endotoxin-degrading bacteria and enzyme compound preparation in Scheme 8 of Example 1 was 4:1.

[0087] The experiment lasted for 4 weeks, with the mice's condition and mortality observed weekly. The Disease Activity Index (DAI) was used to evaluate the mice's health status.

[0088] The DAI scoring principles are as follows: Weight loss <1%, 0 points; ≥1% to <5%, 1 point; ≥5% to <10%, 2 points; ≥10% to <20%, 3 points; ≥20%, 4 points. Fecal occult blood: negative, 0 points; positive, 2 points; visible bleeding, 4 points. Stool consistency: normal, 0 points; loose stool, 2 points; diarrhea, 4 points.

[0089] Four weeks after feeding, blood was collected from the eyeballs. The collected whole blood was allowed to stand at 4°C for 12 hours, centrifuged at 4000 rpm for 20 minutes, and the supernatant was collected. After dilution to an appropriate factor, the serum endotoxin content was determined using the dynamic turbidimetric method. The serum levels of inflammatory factors TNF-α, IL-1β, C-reactive protein (CRP), and lactate dehydrogenase (LDH) were measured using a kit. At the end of the experiment, the mice were anesthetized, euthanized by cervical dislocation, and the abdominal cavity was opened to remove and weigh the organs. The entire colon was removed, its length was recorded, and photographs were taken.

[0090] 1. Disease Activity Index (DAI) in mice

[0091] Table 5. DAI scores of mice in different treatment groups

[0092] Processing group DAI Blank group 0 Model group 6 Pre-degradation test group 7 Degradation test group 2

[0093] The changes in body weight of mice in each group after 28 days are shown in the figure. Figure 2 The DAI scores are shown in Table 5. In the control group, mice exhibited normal activity, thick, smooth, and glossy fur, were responsive, had normal fecal morphology, and were in good condition; no mice died during the 28-day experiment. In the model group, mice experienced significant weight loss after 28 days of feeding, with dry, dull fur, reduced food intake, slow activity, loose stools, small amounts of bloody stools, and lethargy; no mice died during the 28-day experiment. In the pre-degradation experimental group, mice experienced severe weight loss after 28 days of feeding, with dry, dull fur, reduced food intake, slow activity, loose stools, bloody stools, and lethargy; no mice died during the 28-day experiment. Compared to the pre-degradation experimental group, the post-degradation experimental group showed increased weight, improved food intake and mobility, small amounts of loose stools without bloody stools, smooth fur, and good activity; no mice died during the 28-day experiment.

[0094] 2. Endotoxin levels and serum inflammatory factors

[0095] Table 6. Plasma endotoxin levels and serum inflammatory mediator levels in mice under different treatment groups.

[0096]

[0097] Note: Different letters indicate significant differences (p < 0.05) in the same column.

[0098] After 28 days of feeding, the plasma endotoxin levels and serum inflammatory mediator levels in mice are shown in Table 6 below. Compared with the model group and the pre-degradation experimental group, the blood endotoxin levels and serum inflammatory factors TNF-α and IL-1β levels in mice fed the degraded diet were significantly reduced.

[0099] Figure 3 The CRP levels in mice were compared between different treatment groups. In the blank control group, no inflammatory response was observed and CRP was almost not expressed. In the model group, an inflammatory response was observed and CRP expression was increased compared to the blank control group. Before degradation, CRP expression was significantly higher in the experimental group compared to the model group. After degradation, CRP expression was significantly lower in the experimental group compared to both the pre-degradation experimental group and the model group.

[0100] Figure 4 The LDH content of mice in different treatment groups was compared with that in the blank group, the LDH expression of mice in the model group was increased, and the LDH expression of mice in the degraded experimental group was significantly decreased compared with that in the pre-degradation experimental group.

[0101] 3. Organ coefficient

[0102] Table 7 Comparison of organ coefficients in mice from different treatment groups

[0103]

[0104] Note: Different letters indicate significant differences (p < 0.05) in the same column.

[0105] Table 7 shows the comparison of organ coefficients in mice from different treatment groups after 28 days of the experiment. Compared with the pre-degradation group, the spleen and liver coefficients were significantly reduced, and the colon length was increased in the post-degradation group (see Table 7). Figure 5 ).

[0106] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A microecological preparation, characterized in that, The microecological preparation is made from Lactobacillus plantarum CGMCC NO. 12849, Bacillus subtilis CGMCC NO. 25499 and Saccharomyces cerevisiae CGMCC NO. 21678.

2. The microecological preparation according to claim 1, characterized in that, The microecological preparation is made from Lactobacillus plantarum CGMCC NO.12849 bacterial powder, Bacillus subtilis CGMCC NO.25499 bacterial powder, and Saccharomyces cerevisiae CGMCC NO.21678 bacterial powder, and the viable count of the bacterial powder is 1×10⁻⁶. 9 ~1×10 11 CFU / g, preferably, the viable count of the *Lactobacillus plantarum* CGMCC NO.12849 bacterial powder is 1×10⁻⁶. 10 The viable count of the Bacillus subtilis CGMCC NO.25499 bacterial powder was 6 × 10⁻⁶ CFU / g. 9 The viable count of the yeast CGMCC NO.21678 powder was 3 × 10⁻⁶ CFU / g. 9 CFU / g.

3. The microecological preparation according to claim 1, characterized in that, The microecological preparation is made from Lactobacillus plantarum CGMCC NO.12849 bacterial powder, Bacillus subtilis CGMCC NO.25499 bacterial powder and Saccharomyces cerevisiae CGMCC NO.21678 bacterial powder in a mass ratio of (0.5~2):(1~5):(0.5~2), preferably 1:5:

1.

4. A compound preparation containing the microecological preparation according to any one of claims 1 to 3, characterized in that, The compound preparation also includes a compound enzyme preparation, and the mass ratio of the microecological preparation to the compound enzyme preparation is 13:

7.

5. The compound formulation according to claim 4, characterized in that, The compound enzyme preparation includes cellulase, xylanase, and pectinase, wherein the mass ratio of cellulase, xylanase, and pectinase is 5:1:

1.

6. A fermented feed, characterized in that, The fermented feed is produced by fermentation of the compound preparation described in claim 4 or 5.

7. The method for preparing fermented feed according to claim 6, characterized in that, Add 3% to 7% of a compound preparation and 10% molasses to the feed ingredients, control the moisture content of the fermented feed to 50% to 60%, pH 5.5 to 6.0, and culture at 30℃ for 3 to 4 days to obtain the final product.

8. The preparation method according to claim 7, characterized in that, The feed ingredients include Escherichia coli single-cell protein and wheat bran, with the mass ratio of Escherichia coli single-cell protein to wheat bran being 4:

1.

9. The application of the microecological preparation according to any one of claims 1 to 3 or the compound preparation according to claim 4 or 5 in the degradation of endotoxins in feed, wherein the application includes: The preparation is added to the feed through fermentation or non-fermentation methods.

10. The application according to claim 9, characterized in that, Fermenting feed with the microecological preparations described in any one of claims 1 to 3 or the compound preparations described in claims 4 or 5 can reduce the endotoxin content in the feed.

11. The use of the microecological preparation according to any one of claims 1 to 3 or the compound preparation according to claim 4 or 5 in the preparation of feed that improves animal intestinal health.

12. The application according to claim 11, characterized in that, The improvement of gut health includes at least one of the following: reducing serum endotoxins, inhibiting serum inflammatory factors, and improving colon length.

13. The application according to claim 11, characterized in that, The application can also achieve at least one of the following effects: improve growth performance, reduce serum C-reactive protein and lactate dehydrogenase levels, improve spleen coefficient, and improve liver coefficient.

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