Method for improving the ability of broiler chickens to resist lps stress

Abstract

The application provides a method for improving the LPS stress resistance of broilers, which is realized by feeding a mannose oligosaccharide compound preparation prepared from enzymolysis coffee grounds. The preparation method of the mannose oligosaccharide compound preparation comprises the following steps: crushing coffee grounds, and adding a compound reagent for pretreatment; adding a compound enzyme preparation I to the mixed system after reaction, and carrying out enzymolysis for 6-8 hours under the condition of 50-60 DEG C; then adding a compound enzyme preparation II, and carrying out sufficient enzymolysis under the condition of 55-65 DEG C; after the enzymolysis is completed, a fermentation compound microbial agent is used for continuing fermentation for 6-12 hours under the condition of 30-35 DEG C, and drying is carried out; the compound enzyme preparation I comprises papain, alpha-amylase and saccharifying enzyme; the compound enzyme preparation II comprises cellulase, mannanase and glucanase; and the fermentation compound microbial agent comprises maxwell kluyver yeast and lactobacillus plantarum. The method can not only avoid drug residues caused by feeding drugs, but also realize the reuse of coffee grounds.

Description

Technical Field

[0001] This invention belongs to the field of broiler breeding technology, specifically relating to a method for improving the resistance of broilers to LPS stress. Background Technology

[0002] Broiler chickens are susceptible to various stressors during the rearing process. The conventional approach is to use drugs to prevent or treat them after they have been exposed to stressors. However, there is a risk of drug residues after feeding them drugs. If people consume broiler chickens with drug residues, it may harm their health.

[0003] Mannan oligosaccharides, also known as mannan oligosaccharides, are a novel class of antigenically active substances extracted from the cell walls of yeast cultures. They are widely found in konjac flour, guar gum, guar gum, and the cell walls of various microorganisms, and are essentially short-chain carbohydrates. Due to their excellent physicochemical properties, including high-temperature resistance, stability, easy water solubility, and safety and non-toxicity, as well as their ability to protect the intestines and enhance immunity, they are widely used as feed additives in the feed industry both domestically and internationally.

[0004] Coffee grounds are the residue left over from the production of instant coffee; approximately 0.9 tons of coffee grounds are produced for every ton of instant coffee. Reports indicate that coffee grounds contain a large amount of sugars, fats, polyphenols, and minerals. For example, they contain various sugars such as mannan (21.2–57.0 wt%), galactose (11.0–30.4 wt%), glucose (8.6–19.0 ​​wt%), and arabinose (1.7–6.0 wt%). As a byproduct of coffee production, coffee grounds are inexpensive and have low utilization rates.

[0005] Therefore, this invention combines the two to provide a new and healthier method for improving the resistance of broilers to LPS stress. Summary of the Invention

[0006] To address the technical problems existing in the prior art, this invention provides a method for improving the resistance to LPS stress in broilers. This method is achieved by feeding broilers with a mannan oligosaccharide compound preparation made by enzymatic hydrolysis of coffee grounds. This method avoids drug residues caused by conventional drug feeding and enables the high-value reuse of coffee grounds. The method is safe, economical, and pollution-free.

[0007] The present invention adopts the following technical solution:

[0008] A method to improve the resistance of broilers to LPS stress is achieved by feeding them a mannan oligosaccharide compound preparation made from enzymatically hydrolyzed coffee grounds;

[0009] The preparation method of the mannan oligosaccharide complex preparation includes the following steps: pulverizing the collected coffee grounds and adding a complex reagent for pretreatment, wherein the complex reagent is an aqueous solution of sodium carbonate and calcium oxide;

[0010] Add compound enzyme preparation I to the mixed system after the reaction, and enzymatically hydrolyze for 6-8 hours at 50-60℃;

[0011] Then add compound enzyme preparation II and fully enzymatically hydrolyze at 55~65℃;

[0012] After enzymatic hydrolysis, continue fermentation for 6-12 hours at 30-35℃ using a fermentation compound inoculum, and then dry the product after fermentation.

[0013] The compound enzyme preparation I includes papain, α-amylase, and saccharifying enzyme;

[0014] The compound enzyme preparation II includes cellulase, mannanase, and dextranase;

[0015] The fermentation compound microbial agent includes Kluyveromyces macrocephala and Lactobacillus plantarum.

[0016] Furthermore, the concentration of the sodium carbonate aqueous solution in the compound reagent is 3wt%~5wt%, and the concentration of the calcium oxide solution is 1wt%~3wt%, with a volume ratio of sodium carbonate aqueous solution to calcium oxide aqueous solution of 2~3:1; and a mass ratio of coffee grounds to the compound reagent mixture of 1:2~3.

[0017] Furthermore, the mass ratio of papain, α-amylase, and saccharifying enzyme in the above-mentioned compound enzyme preparation I is (2~3):(1~2.5):(0.5~1); wherein the enzyme activity of papain is 2000~30000U / g, the enzyme activity of α-amylase is 6000~20000U / g, and the enzyme activity of saccharifying enzyme is 8000~60000U / g.

[0018] Furthermore, the mass ratio of cellulase, mannanase, and dextranase in compound enzyme preparation II is (1~2):(2~4):(1.5~3); wherein the enzyme activity of cellulase is 8000~30000U / g, the enzyme activity of mannanase is 10000~80000U / g, and the enzyme activity of dextranase is 8000~20000U / g.

[0019] Furthermore, the viable count of *Kluyveromyces martensii* in the fermentation compound inoculant is not less than 7 × 10⁻⁶. 9 CFU / g and viable count of Lactobacillus plantarum not less than 5 × 10⁻⁶ 10 cfu / g.

[0020] Furthermore, the fermentation compound microbial agent is dissolved in brown sugar water, stirred evenly, and then added to the mixed fermentation liquid.

[0021] Furthermore, the concentration of the brown sugar water is 5-10 wt%.

[0022] Furthermore, the drying temperature is 55~65℃.

[0023] Furthermore, the amount of the mannan oligosaccharide compound preparation added is 1-5 wt% of the total basal diet.

[0024] Furthermore, the mannan oligosaccharide complex preparation contains 24.6 wt% mannan oligosaccharide, 12.67 wt% crude protein, 11.03 wt% small peptides, 3.08 wt% lactic acid, and 1.64 wt% acetic acid.

[0025] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0026] This invention established an LPS broiler chicken experimental model, and the prepared mannan oligosaccharide compound preparation improved the growth performance and antioxidant capacity of broilers. This avoids drug residues caused by feeding broilers with drugs to enhance their immunity, and also achieves high-value, pollution-free utilization of coffee grounds.

[0027] This invention pre-treats coffee grounds, then enzymatically hydrolyzes and ferments them, breaking down the large, difficult-to-degrade molecules in the coffee grounds into easily absorbed small molecules, which is more beneficial for broiler chickens to absorb. This method achieves a mannan oligosaccharide conversion rate of 80%-95%.

[0028] Coffee grounds have a high crude fiber content, including 8.6 wt% to 15.3 wt% cellulose and 36.7 wt% hemicellulose. This invention uses a chemical method to pretreat coffee grounds, which changes the original molecular structure of the raw material and alters the chemical bonds (such as hydrogen bonds). As the pretreatment proceeds, many micropores are formed on the fiber surface. These micropores increase the contact area of ​​the substrate and make it easier for the complex enzyme to enter the interior of the substrate, thus improving the enzymatic hydrolysis efficiency.

[0029] Furthermore, compared with conventional NaOH pretreatment, the present invention uses a composite reagent of sodium carbonate and calcium oxide, which is less alkaline than NaOH. At the same time, calcium oxide releases heat when it comes into contact with water, providing some heat for subsequent enzymatic hydrolysis.

[0030] Previous researchers mostly used mannanase, cellulase, etc., without considering the decomposition and utilization of other components of coffee grounds. This invention uses a combination of compound enzyme preparation I and compound enzyme preparation II, which not only solves the problem of the influence of other macromolecules and improves the conversion rate of mannan, but also provides more nutrients for animals.

[0031] The present invention involves a brief fermentation process after enzymatic hydrolysis, which avoids the consumption of oligosaccharides and increases beneficial metabolites, thereby enhancing the reuse value of coffee grounds, a byproduct of coffee production.

[0032] In addition, the Kluwer Saccharomyces roxburghii in the fermentation compound microbial agent was included in the feed and feed additive catalog in July 2023. Compared with other brewing yeasts, it has unique advantages such as high temperature resistance, fast growth rate, high biomass, and fruit wine aroma after fermentation, which can increase animal feed intake and reduce diarrhea rate. Detailed Implementation

[0033] The formulations prepared by the above method are also within the scope of protection of this application.

[0034] The present invention will be further described in detail below with reference to specific embodiments, so that those skilled in the art can more clearly understand the present invention. The following embodiments are only for illustrating the present invention and are not intended to limit the scope of the present invention. Based on the specific embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the protection scope of the present invention.

[0035] In the embodiments of the present invention, unless otherwise specified, all raw material components are commercially available products well known to those skilled in the art; in the embodiments of the present invention, unless specifically specified, the technical means used are conventional means well known to those skilled in the art. In the embodiments of the present invention, all raw materials used are conventional commercially available products.

[0036] Calcium oxide: CAS No. 1305-78-8, CP (Shanghai Test), ≥97.0%.

[0037] A method to improve the resistance of broilers to LPS stress is achieved by feeding them a mannan oligosaccharide compound preparation made from enzymatically hydrolyzed coffee grounds;

[0038] The preparation method of the mannan oligosaccharide complex preparation includes the following steps: pulverizing the collected coffee grounds and adding a complex reagent for pretreatment, wherein the complex reagent is an aqueous solution of sodium carbonate and calcium oxide;

[0039] Add compound enzyme preparation I to the mixed system after the reaction, and enzymatically hydrolyze for 6-8 hours at 50-60℃;

[0040] Then add compound enzyme preparation II and fully enzymatically hydrolyze at 55~65℃;

[0041] After enzymatic hydrolysis, continue fermentation for 6-12 hours at 30-35℃ using a fermentation compound inoculum, and then dry the product after fermentation.

[0042] The compound enzyme preparation I includes papain, α-amylase, and saccharifying enzyme;

[0043] The compound enzyme preparation II includes cellulase, mannanase, and dextranase;

[0044] The fermentation compound microbial agent includes Kluyveromyces macrocephala and Lactobacillus plantarum.

[0045] Furthermore, the concentration of the sodium carbonate aqueous solution in the compound reagent is 3wt%~5wt%, and the concentration of the calcium oxide solution is 1wt%~3wt%, with a volume ratio of sodium carbonate aqueous solution to calcium oxide aqueous solution of 2~3:1; and a mass ratio of coffee grounds to the compound reagent mixture of 1:2~3.

[0046] Furthermore, the mass ratio of papain, α-amylase, and saccharifying enzyme in the above-mentioned compound enzyme preparation I is (2~3):(1~2.5):(0.5~1); wherein the enzyme activity of papain is 2000~30000U / g, the enzyme activity of α-amylase is 6000~20000U / g, and the enzyme activity of saccharifying enzyme is 8000~60000U / g.

[0047] Furthermore, the mass ratio of cellulase, mannanase, and dextranase in compound enzyme preparation II is (1~2):(2~4):(1.5~3); wherein the enzyme activity of cellulase is 8000~30000U / g, the enzyme activity of mannanase is 10000~80000U / g, and the enzyme activity of dextranase is 8000~20000U / g.

[0048] Furthermore, the viable count of *Kluyveromyces martensii* in the fermentation compound inoculant is not less than 7 × 10⁻⁶. 9 CFU / g and viable count of Lactobacillus plantarum not less than 5 × 10⁻⁶ 10 cfu / g.

[0049] Furthermore, the fermentation compound microbial agent is dissolved in brown sugar water, stirred evenly, and then added to the mixed fermentation liquid.

[0050] Furthermore, the concentration of the brown sugar water is 5-10 wt%.

[0051] Furthermore, the drying temperature is 55~65℃.

[0052] Furthermore, the amount of the mannan oligosaccharide compound preparation added is 1-5 wt% of the total basal diet.

[0053] Example 1

[0054] This invention provides a method for improving the resistance to LPS stress in broilers, achieved by feeding them a mannan oligosaccharide complex preparation made from enzymatically hydrolyzed coffee grounds; the preparation method of the mannan oligosaccharide complex preparation includes the following steps:

[0055] (1) Take commercially available coffee grounds and grind them to 40-60 mesh;

[0056] (2) Weigh the pulverized coffee grounds from (1) and add sodium carbonate and calcium oxide solution at a mass ratio of 1:2.5, wherein the concentration of sodium carbonate in the solution is 3wt% and the concentration of calcium oxide is 1wt%, and let stand at room temperature for 4 hours.

[0057] (3) After pretreatment, the mixture from (2) is transferred to a fermenter, and compound enzyme preparation I is added to the fermenter at 4 wt% of the system. Compound enzyme preparation I includes papain, α-amylase, and saccharifying enzyme in a mass ratio of 3:1:1, with papain having an activity of 20,000 U / g, α-amylase having an activity of 12,000 U / g, and saccharifying enzyme having an activity of 40,000 U / g. Enzymatic hydrolysis is performed in a 55℃ fermenter with continuous stirring for 8 hours.

[0058] (4) After the enzymatic hydrolysis is completed, add compound enzyme preparation II to the system in (3) to continue the enzymatic hydrolysis reaction. The amount added is 10 wt% of the system. Compound enzyme preparation II includes cellulase, mannanase, and dextranase in a mass ratio of 1:3:1.5. The cellulase activity is 28000 U / g, the mannanase activity is 50000 U / g, and the dextranase activity is 15000 U / g. The enzymatic hydrolysis conditions are a 60℃ fermenter with continuous stirring for 24 hours.

[0059] (4) After the enzymatic hydrolysis is completed, the fermenter is cooled down to 32°C.

[0060] (5) Prepare a 5wt% concentration of brown sugar water, add the fermentation compound microbial agent to the brown sugar water and dissolve it. The amount of the compound fermentation microbial agent added is 0.5wt% of the system. The fermentation compound microbial agent contains Kluyveromyces martensii and Lactobacillus plantarum. The viable count of Kluyveromyces martensii is 7×10⁻⁶. 9 The cfu / g and viable count of Lactobacillus plantarum were 2×10⁻⁶. 10 The concentration of cfu / g was 2:1 by mass. The fermentation conditions were 32℃ for 6 hours.

[0061] (6) After fermentation, place the mixture in a 60°C oven to dry, and you will get the mannan oligosaccharide complex preparation.

[0062] The detection methods are as follows:

[0063] Moisture content testing method: GB / T 6435-2014

[0064] Crude protein detection method: GB / T 6432-2018

[0065] Method for detecting acid-soluble proteins: NY / T 3801-2020

[0066] Organic acid detection method: GB / T 23877-2009

[0067] Mannan oligosaccharide detection method: QB / T yeast β-glucan

[0068] Table 1. Results of indicator testing

[0069]

[0070] The coffee grounds treated using the method of this invention have a mannan conversion rate of 80%-95%.

[0071] The formula for calculating the mannan conversion rate is: mannan oligosaccharide content / raw material mannan content. 100%.

[0072] Example 2

[0073] In this embodiment, the mannan oligosaccharide prepared in Example 1 was used in a broiler LPS model experiment to explore its antioxidant activity. The specific method is as follows:

[0074] Broilers of similar weight and in good health were selected and divided into three treatment groups based on the principle of equal distribution of males and females of similar weight. Each treatment group had four replicates, with 14 birds per replicate. The indoor environment was well-ventilated and the room temperature was maintained at 22-25℃. The laying hens were floor-raised and had free access to feed and water. The animals' mental state, feed and water intake, and fecal color were observed daily. Daily mortality and morbidity were recorded.

[0075] Control group 1: Basal diet;

[0076] Control group 2-LPS group: basal diet + LPS stress;

[0077] Experimental Group 1 - LPS-compound preparation group: basal diet + LPS stress + 2wt% mannan oligosaccharide compound preparation prepared in Example 1.

[0078] The basal diet consists of 62.5% corn, 24% soybean meal, 2% rice bran, 0.5% soybean oil, 8% limestone powder, and 3% premix by weight.

[0079] In control group 1, each chicken was injected intraperitoneally with physiological saline, while the other groups were injected intraperitoneally with LPS. After feeding, the growth performance of the broilers in each group was observed and recorded, and the impact on the broiler's immune performance was analyzed.

[0080] Table 2 Effects of mannan oligosaccharide compound preparations on broiler growth performance

[0081]

[0082] As shown in Table 2, compared with control group 1, control group 2 had a lower average daily weight gain, and higher feed conversion ratio and mortality rate. Compared with control group 1, the growth performance of broilers in the basal diet + LPS stress + 2wt% mannan oligosaccharide group prepared in Example 1 was increased to varying degrees.

[0083] Table 3. Effects of mannan oligosaccharide compound preparation on serum immune indicators in broilers.

[0084]

[0085] The increased levels of IgM, IgA, IgG, and SOD in experimental group 1 indicate that the mannan oligosaccharide prepared from coffee grounds has a positive effect on the immune performance of laying hens.

[0086] Comparative Example 1

[0087] This comparative example is used to explore the effect of the preparation method on the composition of the mannan oligosaccharide complex formulation. It differs from Example 1 in that fermentation was not performed after enzymatic hydrolysis. Specifically, it includes the following steps:

[0088] (1) Take commercially available coffee grounds and grind them to 40-60 mesh;

[0089] (2) Weigh the pulverized coffee grounds from (1) and add sodium carbonate and calcium oxide solution at a mass ratio of 1:2.5, wherein the concentration of sodium carbonate in the solution is 3wt% and the concentration of calcium oxide is 1wt%, and let stand at room temperature for 4 hours.

[0090] (3) After pretreatment, the mixture from (2) is transferred to a fermenter, and compound enzyme preparation I is added to the fermenter at 4 wt% of the system. Compound enzyme preparation I includes papain, α-amylase, and saccharifying enzyme in a mass ratio of 3:1:1, with papain having an activity of 20,000 U / g, α-amylase having an activity of 12,000 U / g, and saccharifying enzyme having an activity of 40,000 U / g. Enzymatic hydrolysis is performed in a 55℃ fermenter with continuous stirring for 8 hours.

[0091] (4) After the enzymatic hydrolysis is completed, add compound enzyme preparation II to the system in (3) to continue the enzymatic hydrolysis reaction. The amount added is 10 wt% of the system. Compound enzyme preparation II includes cellulase, mannanase, and dextranase in a mass ratio of 1:3:1.5. The cellulase activity is 28000 U / g, the mannanase activity is 50000 U / g, and the dextranase activity is 15000 U / g. The enzymatic hydrolysis conditions are a 60℃ fermenter with continuous stirring for 24 hours.

[0092] (5) After the enzymatic hydrolysis is completed, place it in a 60℃ oven to dry, and the finished product can be obtained.

[0093] Table 4 Indicator Detection Results

[0094]

[0095] Table 5. Effects of the formulation on growth performance and serum immune indicators in broilers.

[0096]

[0097] In Comparative Example 1, the immune indicators were all lower than those in Example 1, and the material weight ratio was higher than that in Example 1.

[0098] Comparative Example 2

[0099] This comparative example is used to explore the effect of preparation method on the composition of mannan oligosaccharide complex formulation. It differs from Example 1 in that only complex enzyme preparation II was added during enzymatic hydrolysis, without the addition of complex enzyme preparation I. The specific steps include:

[0100] (1) Take commercially available coffee grounds and grind them to 40-60 mesh;

[0101] (2) Weigh the pulverized coffee grounds from (1) and add sodium carbonate and calcium oxide solution at a mass ratio of 1:2.5, wherein the concentration of sodium carbonate in the solution is 3wt% and the concentration of calcium oxide is 1wt%, and let stand at room temperature for 4 hours.

[0102] (3) After pretreatment, a compound enzyme was added to the system in (2) for enzymatic hydrolysis. The amount added was 10 wt% of the system. The compound enzyme preparation included cellulase, mannanase, and dextranase in a mass ratio of 1:3:1.5. The cellulase activity was 28,000 U / g, the mannanase activity was 50,000 U / g, and the dextranase activity was 15,000 U / g. The enzymatic hydrolysis conditions were a 60℃ fermenter with continuous stirring for 24 hours.

[0103] (4) After the enzymatic hydrolysis is completed, the fermenter is cooled down to 32°C.

[0104] (5) Prepare a 5wt% concentration of brown sugar water, add the fermentation compound microbial agent to the brown sugar water and dissolve it. The amount of the compound fermentation microbial agent added is 0.5wt% of the system. The fermentation compound microbial agent contains Kluyveromyces martensii and Lactobacillus plantarum. The viable count of Kluyveromyces martensii is 7×10⁻⁶. 9 The cfu / g and viable count of Lactobacillus plantarum were 2×10⁻⁶. 10 The concentration of cfu / g was 2:1 by mass. The fermentation conditions were 32℃ for 6 hours.

[0105] (6) After fermentation, place the product in a 60℃ oven to dry, and the finished product can be obtained.

[0106] Table 6 Indicator Detection Results

[0107]

[0108] Table 7 Effects of the formulation on growth performance and serum immune indicators in broilers

[0109]

[0110] In Comparative Example 2, the immune indicators were all lower than those in Example 1.

[0111] Comparative Example 3

[0112] This comparative example was used to investigate the effect of replacing papain in compound enzyme preparation I with neutral protease to obtain a new compound enzyme preparation, named compound enzyme preparation III, on the components of the mannan oligosaccharide compound preparation; at the same time, it was fed to broilers to investigate the effect on the broilers' resistance to LPS stress.

[0113] (1) Take commercially available coffee grounds and grind them to 40-60 mesh;

[0114] (2) Weigh the pulverized coffee grounds from (1) and add sodium carbonate and calcium oxide solution at a mass ratio of 1:2.5, wherein the concentration of sodium carbonate in the solution is 3wt% and the concentration of calcium oxide is 1wt%, and let stand at room temperature for 4 hours.

[0115] (3) After pretreatment, the mixture from (2) is transferred to a fermenter, and compound enzyme preparation III is added to the fermenter at 4 wt% of the system. Compound enzyme preparation I includes neutral protease, α-amylase, and saccharifying enzyme in a mass ratio of 3:1:1, with the neutral protease having an activity of 25000 U / g, the α-amylase having an activity of 12000 U / g, and the saccharifying enzyme having an activity of 40000 U / g. The enzymatic hydrolysis is performed in a 55℃ fermenter with continuous stirring for 8 hours.

[0116] (4) After the enzymatic hydrolysis is completed, add compound enzyme preparation II to the system in (3) to continue the enzymatic hydrolysis reaction. The amount added is 10 wt% of the system. Compound enzyme preparation II includes cellulase, mannanase, and dextranase in a mass ratio of 1:3:1.5. The cellulase activity is 28000 U / g, the mannanase activity is 50000 U / g, and the dextranase activity is 15000 U / g. The enzymatic hydrolysis conditions are a 60℃ fermenter with continuous stirring for 24 hours.

[0117] (4) After the enzymatic hydrolysis is completed, the fermenter is cooled down to 32°C.

[0118] (5) Prepare a 5wt% concentration of brown sugar water, add the fermentation compound microbial agent to the brown sugar water and dissolve it. The amount of the compound fermentation microbial agent added is 0.5wt% of the system. The fermentation compound microbial agent contains Kluyveromyces martensii and Lactobacillus plantarum. The viable count of Kluyveromyces martensii is 7×10⁻⁶.9 The cfu / g and viable count of Lactobacillus plantarum were 2×10⁻⁶. 10 The concentration of cfu / g was 2:1 by mass. The fermentation conditions were 32℃ for 6 hours.

[0119] (6) After fermentation, place the mixture in a 60°C oven to dry, and you will get the mannan oligosaccharide compound preparation. The same method was used for broiler chicken breeding trials.

[0120] Table 8 Effects of the formulation on broiler growth performance and serum immune indicators

[0121]

[0122] In Comparative Example 3, the immune indicators were all lower than those in Example 1, and the difference in feed conversion ratio was more obvious. This is because papain is a protease that can decompose proteins in acidic, neutral, and alkaline environments, and can reduce the bitterness in feed.

[0123] It should be noted that the above embodiments are only for further elaboration and explanation of the technical solution of the present invention, and are not intended to further limit the technical solution of the present invention. The method of the present invention is only a preferred embodiment and is not intended to limit the scope of protection of the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A method for improving the ability of broiler chickens to resist LPS stress, characterized in that, The preparation of the mannose oligosaccharide complex preparation is achieved by feeding enzymatic hydrolysis of coffee grounds; The preparation method of the mannose oligosaccharide complex preparation comprises the following steps: the collected coffee grounds are subjected to crushing treatment, and a composite reagent is added for pretreatment, wherein the composite reagent is an aqueous solution of sodium carbonate and calcium oxide; After the reaction, composite enzyme preparation I is added to the mixed system, and enzymatic hydrolysis is carried out at 50-60 DEG C for 6-8 hours; Then, composite enzyme preparation II is added, and sufficient enzymatic hydrolysis is carried out at 55-65 DEG C. After the enzymatic hydrolysis is completed, fermentation is continued for 6-12 hours at 30-35 DEG C using a fermentation composite microbial agent, and then drying treatment is carried out. The composite enzyme preparation I comprises papain, alpha-amylase and saccharifying enzyme. The composite enzyme preparation II comprises cellulase, mannanase and glucanase. The fermentation composite microbial agent comprises Kluyveromyces marxianus and Lactobacillus plantarum. The concentration of the aqueous sodium carbonate solution in the composite reagent is 3wt%-5wt%, the concentration of the solution prepared from calcium oxide is 1wt%-3wt%, and the volume ratio of the aqueous sodium carbonate solution to the aqueous calcium oxide solution is 2-3:1; and the mass ratio of the coffee grounds to the composite reagent mixture is 1:2-3. In the composite enzyme preparation I, the mass ratio of papain, alpha-amylase and saccharifying enzyme is (2-3):(1-2.5):(0.5-1); the enzyme activity of papain is 2000-30000 U / g, the enzyme activity of alpha-amylase is 6000-20000 U / g, and the enzyme activity of saccharifying enzyme is 8000-60000 U / g. In the composite enzyme preparation II, the mass ratio of cellulase, mannanase and glucanase is (1-2):(2-4):(1.5-3); the enzyme activity of cellulase is 8000-30000 U / g, the enzyme activity of mannanase is 10000-80000 U / g, and the enzyme activity of glucanase is 8000-20000 U / g.

2. The method for improving the LPS stress resistance of broilers according to claim 1, characterized in that, The viable cell count of K. marxianus in the fermentation complex microbial agent is not less than 7 x 10 9 The viable cell count of L. plantarum is not less than 5 x 10 10 cfu / g.

3. The method for improving the LPS stress resistance of broilers according to claim 1, characterized in that, The fermentation composite microbial agent is dissolved in brown sugar water, and then added into the mixed fermentation liquid after stirring.

4. The method for improving the LPS stress resistance of broilers according to claim 3, characterized in that, The concentration of the brown sugar water is 5wt%-10wt%.

5. The method for improving the LPS stress resistance of broilers according to claim 1, characterized in that, The drying temperature is 55-65 DEG C.

6. The method for improving the LPS stress resistance of broilers according to claim 1, characterized in that, The addition amount of the mannose oligosaccharide complex preparation is 1wt%-5wt% of the total amount of the basic daily ration.

7. The method for improving the LPS stress resistance of broilers according to claim 1, wherein, The mannose oligosaccharide complex preparation contains mannose oligosaccharide 24.6wt%, crude protein 12.67wt%, small peptides 11.03wt%, lactic acid 3.08wt% and acetic acid 1.64wt%.

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