A high-temperature-resistant composite microbial granular preparation, a processing method thereof and application thereof

Abstract

The present application relates to the technical field of microbial feed additives, and discloses a high-temperature-resistant composite microbial granular preparation and a processing method and application thereof, comprising the following steps: in a low-temperature sterile environment, a trehalose protective agent, a vitamin E stabilizer and a natural plant colorant are sequentially added into purified water and stirred; modified corn starch and defatted rice bran are uniformly mixed in proportion to obtain a composite porous carrier; Bacillus coagulans live powder, Bacillus licheniformis live powder and Clostridium butyricum live powder are uniformly scattered into the composite porous carrier, a protective colorant liquid is atomized and sprayed, and stirring and mixing are performed; the mixed material is subjected to high-temperature forming through a double-screw extrusion process to obtain primary formed granules; the primary formed granules are subjected to constant-temperature air drying, natural cooling and screening, and the finished granules with uniform specifications are selected and stored in a light-proof and sealed manner. The present application has the advantages of green safety, strong live bacteria stability, large-scale production suitability and high feeding precision, and can be widely applied to the field of standard ecological pig breeding.

Description

Technical Field

[0001] This invention relates to the field of microbial feed additive technology, and in particular to a high-temperature resistant composite microbial granule preparation, its processing method, and its application. Background Technology

[0002] Under large-scale, high-density pig farming, problems such as stress and gut microbiota imbalance frequently occur, easily causing damage to the intestinal mucosa and digestive disorders, leading to symptoms such as diarrhea and decreased immunity. This directly results in low feed utilization and high feed conversion ratios. Simultaneously, pig excrement contains excessive levels of ammonia nitrogen and hydrogen sulfide, and the pigpens suffer from severe odor pollution, hindering the development of ecological and standardized pig farming. Probiotics, as residue-free, green, and safe feed additives, are core products for replacing antibiotic-based farming and improving pig performance.

[0003] Currently available commercially available compound probiotic preparations for pigs and related patented technologies generally suffer from four core technical defects, severely hindering industrialization and promotion: First, their high-temperature resistance is extremely poor. The 70-80℃ extrusion granulation process commonly used in feed industrial production leads to the inactivation of a large number of live probiotics. The survival rate of live bacteria after high-temperature granulation in existing products is generally less than 30%, resulting in a significant reduction in their effectiveness. Second, the scientific formulation of strains is insufficient. Most products are simply mixtures of strains, failing to achieve functional complementarity and synergistic ratios without antagonism, and thus cannot simultaneously achieve multiple effects such as intestinal antibacterial activity, mucosal repair, and feed enhancement. Third, the products have low standardization. They are mostly powders or irregular granules, leading to problems such as dust loss, uneven mixing, and difficulty in accurately controlling the dosage. There are no standardized size parameters or visual labels. Fourth, their process adaptability is poor. Most technologies are only suitable for room-temperature mixing scenarios and have not designed a dedicated protection system for high-temperature granulation processes, making mass production difficult.

[0004] Currently, no publicly available literature or patents simultaneously achieve "high-temperature, high-viability bacteria retention at 70-80℃, precise synergistic compounding of three strains, standardized colored granule molding, and large-scale, low-cost mass production" of a probiotic preparation specifically for pigs. Addressing the numerous shortcomings of existing technologies, this invention specifically develops a high-temperature resistant, highly active, standardized, and multifunctional composite microbial granule preparation to solve common technical challenges in the industry.

[0005] Therefore, those skilled in the art urgently need to develop a high-temperature resistant composite microbial particle formulation, its processing method, and its application. Summary of the Invention

[0006] The purpose of this invention is to overcome the technical defects of existing probiotic feed additives, such as poor heat resistance, severe attenuation of live bacteria during high-temperature granulation, weak synergy of strains, low product standardization, poor feed adaptability, and single function, and to provide a high-temperature resistant compound microbial granule preparation, its processing method, and its application.

[0007] To achieve the above objectives, the present invention provides a high-temperature resistant composite microbial granule preparation, comprising the following raw materials in parts by weight: 2-8 parts of Bacillus coagulans live bacteria powder, 1-5 parts of Bacillus licheniformis live bacteria powder, 0.6-3 parts of Clostridium butyricum live bacteria powder, 60-80 parts of modified corn starch, 10-20 parts of defatted rice bran, 2-5 parts of trehalose protectant, 0.2-0.8 parts of vitamin E stabilizer, 0.1-0.5 parts of natural plant colorant, and 5-10 parts of purified water.

[0008] Furthermore, the modified corn starch is hydroxypropyl modified corn starch, which has high temperature stability at 70-80℃ and a porous encapsulation structure; the trehalose and vitamin E constitute a composite high temperature protection system to achieve high activity retention of the strain under high temperature granulation; the high temperature resistant composite microbial granule preparation is prepared by twin-screw extrusion process, and its particle diameter is 2.0-4.5mm and particle length is 4-13mm.

[0009] Preferably, the optimal raw material composition, by mass parts, includes: 5 parts of Bacillus coagulans live bacteria powder, 3 parts of Bacillus licheniformis live bacteria powder, 1.8 parts of Clostridium butyricum live bacteria powder, 70 parts of modified corn starch carrier, 15 parts of defatted rice bran, 3.5 parts of trehalose protectant, 0.5 parts of vitamin E stabilizer, 0.3 parts of natural plant colorant, and 7.5 parts of purified water.

[0010] This optimal formulation can achieve a survival rate of ≥97.9% for live bacteria under high-temperature granulation at 75℃, and has the best overall effect on intestinal conditioning, reducing feed conversion ratio, and improving the breeding environment.

[0011] Furthermore, the specific parameters and sources of each raw material are as follows: (1) Bacillus coagulans live powder: Bacillus coagulans type I feed additive produced by Hubei Huayang Technology Development Co., Ltd., in accordance with standard Q / HYKJ04-2021, with an effective live count ≥1.0×10⁻⁶. 10 CFU / g, moisture content ≤10%, powder product, ≥90% pass rate on 0.425mm test sieve, with the characteristics of high temperature resistance and inhibition of the proliferation of harmful intestinal bacteria.

[0012] (2) Bacillus licheniformis live bacteria powder: Bacillus licheniformis feed additive produced by Guangxi Academy of Sciences Institute of Biology Co., Ltd., in accordance with standard Q / GXKXYM001-2026, with an effective live bacteria count ≥1.0×10⁻⁶. 9 CFU / g, moisture content ≤9%, powder product, can efficiently decompose large organic molecules in feed and improve feed digestibility and utilization.

[0013] (3) Clostridium butyricum live bacteria powder: Clostridium butyricum feed additive produced by Inner Mongolia Ketuo Microecological Technology Development Co., Ltd., in accordance with standard Q / NMKT023-2023, with an effective live bacteria count ≥1.0×10 8 CFU / g, moisture content ≤7%, powder product, with the function of repairing the intestinal mucosa of pigs and regulating intestinal metabolism.

[0014] (4) Modified corn starch carrier: Hydroxypropyl modified corn starch, produced by Shandong Fuyang Biotechnology Co., Ltd., in accordance with standard Q / 371426FFS002-2023, with a degree of hydroxypropyl substitution of 0.015-0.020 and a moisture content of ≤14%; This carrier has strong high-temperature adhesion and does not soften or decompose under high-temperature conditions of 70-80℃. Its unique porous structure can effectively encapsulate the strains, isolate them from high-temperature damage, and greatly improve the high-temperature survival rate of the strains.

[0015] (5) The defatted rice bran is a feed ingredient produced by Yihai Kerry (Shanghai) International Trading Co., Ltd., which complies with the standard Q / YHJL0502G-2025. It is degreased at low temperature, with a crude protein content of ≥14%, crude fiber of ≤12%, and moisture content of ≤13%. It can be used as an auxiliary carrier and nutritional supplement to improve the stability of pellet formation and supplement natural nutrition for pigs.

[0016] (6) Trehalose protectant: Crystalline trehalose produced by Jiangsu Aogu Biotechnology Co., Ltd., conforms to standard GB / T23529-2009, is food grade, has a content of ≥98.0%, and has strong high temperature stress protection activity.

[0017] (7) Vitamin E stabilizer: DL-α-tocopherol acetate powder, a feed additive produced by Zhejiang Medicine Co., Ltd., complies with standard Q / SCH045-2025, with a purity of ≥50%, which can inhibit the oxidative inactivation of strains and improve high-temperature stability.

[0018] (8) Natural plant coloring agent: Gardenia yellow natural pigment produced by Guangzhou Tianxu Food Additives Co., Ltd., with a color value of ≥80 and in accordance with standard GB26687.

[0019] This invention also provides a method for processing a high-temperature resistant composite microbial granule preparation, comprising the following steps: S1. Preparation of protective coloring solution: Under low temperature and sterile conditions, trehalose protective agent, vitamin E stabilizer and natural plant coloring agent are added to purified water in sequence, and stirred until the system is homogeneous and transparent. Let it stand for later use. S2. Preparation of composite porous carrier: Modified corn starch and defatted rice bran are mixed evenly in proportion to obtain composite porous carrier; S3. Inoculum embedding and mixing: Bacillus coagulans live powder, Bacillus licheniformis live powder and Clostridium butyricum live powder are evenly sprinkled into the composite porous carrier, protective coloring liquid is sprayed by atomization, and the inoculum is completely embedded by constant temperature stirring to obtain a moist mixture. S4. High-temperature extrusion granulation: The mixture is formed at high temperature through a twin-screw extrusion process to obtain pre-formed granules; S5. Low-temperature air drying and shaping: The initially shaped particles are air-dried at a constant temperature and cooled naturally to obtain composite microbial particles with stable properties. S6. Screening and Packaging: Screening removes unqualified particles, and selects uniform finished particles for storage in a light-proof and sealed manner.

[0020] Furthermore, the mixing ratio of each component by mass is as follows: 2-8 parts of Bacillus coagulans live bacteria powder, 1-5 parts of Bacillus licheniformis live bacteria powder, 0.6-3 parts of Clostridium butyricum live bacteria powder, 60-80 parts of modified corn starch, 10-20 parts of defatted rice bran, 2-5 parts of trehalose protectant, 0.2-0.8 parts of vitamin E stabilizer, 0.1-0.5 parts of natural plant colorant, and 5-10 parts of purified water.

[0021] Furthermore, in step S1, the preparation environment is a dust-free and sterile environment with an ambient temperature of 25-30℃, the stirring time is 10-15 minutes, and the materials are completely dissolved and then left to stand at low temperature for later use; in step S2, a horizontal mixer is used to stir at a uniform speed for 5 minutes to achieve uniform mixing of modified corn starch and defatted rice bran.

[0022] Furthermore, in step S3, the bacteria are first stirred for 8 minutes to achieve initial dispersion, and then the protective coloring liquid is sprayed by atomization. Stirring is continued for 15-20 minutes, and the temperature of the system is controlled at 33-35℃ throughout the process.

[0023] Furthermore, in step S4, the granulation temperature is 70-80℃, the screw speed is 200-220r / min, and a die with a diameter of 2.0-4.5mm is used, matched with a cutter spacing of 4-13mm for extrusion molding.

[0024] Furthermore, in step S5, the air-drying temperature is 40-45℃, the air-drying time is 20-25 minutes, and after removing the free moisture on the surface of the particles, they are allowed to cool naturally to room temperature.

[0025] This invention also provides an application of a high-temperature resistant composite microbial granule preparation, which is directly mixed with pig feed and fed to piglets and growing-finishing pigs at a dosage of 0.2%-0.5% of the total feed mass. It can regulate the balance of intestinal flora in pigs, repair the intestinal mucosa, reduce the diarrhea rate of pigs, optimize feed utilization, reduce the feed conversion ratio, and at the same time reduce the emission of ammonia nitrogen and hydrogen sulfide in pig excrement, thereby improving the pig farming environment.

[0026] Compared with the prior art, the beneficial effects of the present invention are as follows: I. Constructing a Triple Synergistic High-Temperature Protection System to Completely Solve the Industry Pain Point of Probiotic Inactivation During High-Temperature Granulation. Addressing the core shortcomings of traditional probiotic preparations—lack of specific high-temperature protection, easy inactivation during granulation above 70℃, and extremely low survival rates—this invention innovatively constructs a triple synergistic high-temperature protection system: "trehalose water-locking protective film + vitamin E antioxidant + porous modified starch physical encapsulation." This system forms a dense and stable composite protective structure on the bacterial surface, effectively resisting thermal damage, oxidative damage, and structural destruction during high-temperature extrusion granulation at 70-80℃, ensuring the integrity of the bacterial structure and stable metabolic activity. Testing shows that the total viable bacterial survival rate of the preparation after high-temperature granulation is consistently ≥95.5%, more than three times higher than existing conventional products, completely overcoming the industry problem of probiotic inactivation under high-temperature feed granulation processes. Simultaneously, the product exhibits excellent stability during room temperature storage, with a shelf life of up to 18 months, far superior to ordinary probiotic preparations on the market, significantly improving the stability of product storage, transportation, and use.

[0027] II. Precise compounding of two- and three-component bacterial strains achieves multifunctional synergistic effects, overcoming the limitations of single-strain applications. This invention screens a three-component compound bacterial system of Bacillus coagulans, Bacillus licheniformis, and Clostridium butyricum with no antagonism and highly complementary functions. Through extensive experiments, the optimal compounding ratio is optimized to achieve precise functional division and synergistic effects. Among them, Bacillus coagulans targets and inhibits harmful bacteria in the pig's intestines, regulating the intestinal microecological balance; Bacillus licheniformis can efficiently degrade difficult-to-digest macromolecules such as starch and protein in feed, improving feed utilization efficiency; and Clostridium butyricum specifically repairs the intestinal mucosal barrier, enhancing intestinal resistance. The combination of the three forms a four-core function of "bacterial inhibition and pest control, mucosal repair, nutritional enhancement, and pollution reduction and emission reduction," completely overcoming the technical shortcomings of single-strain and conventional simple compound products with limited functions and weak conditioning effects, and comprehensively meeting the needs of pig intestinal health conditioning.

[0028] III. Precisely regulates intestinal health, significantly reducing the incidence of diarrhea in pigs. This invention's proprietary ternary microbial strain system effectively optimizes the intestinal flora structure of pigs, significantly increasing the abundance of beneficial bacteria by over 38%, while simultaneously inhibiting the proliferation of harmful bacteria and reducing their abundance by over 41%. This fundamentally balances the intestinal microecological environment and repairs damaged intestinal mucosal barriers. Large-scale breeding trials have verified that feeding pigs with this invention's compound microbial granule preparation reduced the diarrhea rate by over 50%, significantly decreasing the occurrence of intestinal diseases, improving pig survival rates and overall health, reducing disease prevention and medication costs, and achieving healthy pig farming.

[0029] IV. Optimizing feed utilization and significantly improving the economic benefits of pig farming. Leveraging the highly efficient degradation function of Bacillus licheniformis, combined with the intestinal metabolic regulation effects of Bacillus coagulans and Clostridium butyricum, this invention effectively breaks down anti-nutritional factors in feed, promotes the decomposition and absorption of macromolecular nutrients, and significantly improves the digestibility and absorption rate of feed nutrients in pigs. Farming trial data shows that this formulation can reduce the feed conversion ratio of pigs by 7.98%-11.79%. Without requiring additional farming inputs, it effectively improves pig weight gain efficiency, shortens the farming cycle, significantly reduces feed costs, and substantially enhances the overall economic benefits of large-scale pig farming.

[0030] V. Regulating Body Metabolism for Green Pollution Reduction. The compound microbial strain of this invention can precisely regulate the nitrogen and sulfur metabolic pathways in the pig's intestines, effectively reducing the generation and emission of malodorous and harmful gases such as ammonia nitrogen and hydrogen sulfide in livestock excrement. The reduction in harmful gas emissions can reach over 35%, significantly reducing odor in pigpens, reducing pollution in the farming environment, and improving the pig farming environment. This characteristic perfectly aligns with the national industrial policy of green and ecological livestock farming and can be widely applied to standardized and ecological pig farming systems, contributing to the green and low-carbon upgrading of the livestock industry.

[0031] VI. Standardized and Visualized Pellet Design: Convenient Feeding, Wide Adaptability, and High Practicality. This invention uses customized molds to precisely define pellet size, length, and other forming parameters, producing standardized pellets that are highly compatible with pigs' feeding habits. The product achieves a feed mixing uniformity of over 93.5%, offering advantages such as no dust, no loss, and no grading. Simultaneously, it innovatively adds high-temperature resistant natural plant colorants, enabling visual differentiation of pellets and precise control of feed addition ratios. This completely solves the industry problems of large addition errors, uneven mixing, and poor adaptability of existing probiotic products. The product requires no secondary processing and can be directly mixed with compound feed, fully adaptable to all growth stages of piglets and growing-finishing pigs, perfectly suited for large-scale, standardized pig farming scenarios, and possessing extremely high practicality and versatility.

[0032] VII. Integrated and Simplified Preparation Process, Adaptable to Large-Scale Mass Production. This invention integrates low-temperature encapsulation pretreatment, high-temperature extrusion molding, and low-temperature air drying and shaping into a streamlined process. It requires no high-end specialized production equipment and can be scaled up using existing conventional feed pelleting production lines. The overall production process is simple, easy to operate, has low energy consumption, and controllable production costs. Compared to existing low-temperature secondary processing of probiotic preparations, which involves complex procedures and high mass production costs, this invention has stronger industrial adaptability, lower implementation difficulty, and extremely high industrial mass production capacity and market promotion value. Detailed Implementation

[0033] The present invention will be further described below with reference to specific embodiments. The illustrative embodiments and descriptions herein are used to explain the present invention, but are not intended to limit the present invention.

[0034] Example 1 A high-temperature resistant composite microbial granule preparation, by weight, comprises the following raw material components: 2 parts of Bacillus coagulans live bacteria powder, 1 part of Bacillus licheniformis live bacteria powder, 0.6 parts of Clostridium butyricum live bacteria powder, 60 parts of modified corn starch carrier, 10 parts of defatted rice bran, 2 parts of trehalose protectant, 0.2 parts of vitamin E stabilizer, 0.1 parts of natural plant colorant, and 5 parts of purified water.

[0035] Preparation process: S1. Under a sterile environment at 25℃, add 2 parts trehalose, 0.2 parts vitamin E, and 0.1 parts gardenia yellow pigment to 5 parts purified water, and stir at low speed for 15 minutes until completely dissolved to prepare a protective coloring solution. S2. Mix 60 parts of modified corn starch and 10 parts of defatted rice bran and stir for 5 minutes to obtain a composite carrier; S3. Add 2 parts of Bacillus coagulans, 1 part of Bacillus licheniformis, and 0.6 parts of Clostridium butyricum to the composite carrier, stir for 8 minutes, slowly spray the protective coloring solution, and continue stirring for 15 minutes. The temperature is 35℃ throughout the process. S4. Feed the pellets into a twin-screw extruder, set the pelleting temperature to 70℃ and the screw speed to 200r / min, and use a 2.0mm diameter die and a 4mm cutter spacing to extrude and form pellets with a diameter of 2.0mm and a length of 4mm. S5, air-dry at 40℃ for 25 minutes, cool, sieve, and package to obtain a uniform yellow composite microbial granule preparation.

[0036] Example 2 A high-temperature resistant composite microbial granule preparation, by weight, comprises the following raw material components: 5 parts of Bacillus coagulans live bacteria powder, 3 parts of Bacillus licheniformis live bacteria powder, 1.8 parts of Clostridium butyricum live bacteria powder, 70 parts of modified corn starch carrier, 15 parts of defatted rice bran, 3.5 parts of trehalose protectant, 0.5 parts of vitamin E stabilizer, 0.3 parts of natural plant colorant, and 7.5 parts of purified water.

[0037] Preparation process: S1. Under sterile conditions at 27.5℃, add 3.5 parts trehalose, 0.5 parts vitamin E, and 0.3 parts gardenia yellow pigment to 7.5 parts purified water, and stir at low speed for 12.5 minutes until completely dissolved to prepare a protective coloring solution. S2. Mix 70 parts of modified corn starch and 15 parts of defatted rice bran and stir for 5 minutes to obtain a composite carrier; S3. Add 5 parts of Bacillus coagulans, 3 parts of Bacillus licheniformis, and 1.8 parts of Clostridium butyricum to the composite carrier, stir for 8 minutes, slowly spray the protective coloring solution, and continue stirring for 17.5 minutes. The temperature throughout the process is 34℃. S4. Feed the pellets into a twin-screw extruder, set the pelleting temperature to 75℃ and the screw speed to 210r / min, and use a 3.0mm diameter die and an 8mm cutter spacing to extrude and form pellets with a diameter of 3.0mm and a length of 8mm. S5, air-dry at 42.5℃ for 22.5 min, cool, sieve, and package to obtain a uniform bright yellow composite microbial granule preparation.

[0038] Example 3 A high-temperature resistant composite microbial granule preparation, by weight, comprises the following raw material components: 8 parts of Bacillus coagulans live bacteria powder, 5 parts of Bacillus licheniformis live bacteria powder, 3 parts of Clostridium butyricum live bacteria powder, 80 parts of modified corn starch carrier, 20 parts of defatted rice bran, 5 parts of trehalose protectant, 0.8 parts of vitamin E stabilizer, 0.5 parts of natural plant colorant, and 10 parts of purified water.

[0039] Preparation process: S1. Under a sterile environment at 30℃, add 5 parts trehalose, 0.8 parts vitamin E, and 0.5 parts gardenia yellow pigment to 10 parts purified water, and stir at low speed for 10 minutes until completely dissolved to prepare a protective coloring solution. S2. Mix 80 parts of modified corn starch and 20 parts of defatted rice bran and stir for 5 minutes to obtain a composite carrier; S3. Add 8 parts of Bacillus coagulans, 5 parts of Bacillus licheniformis, and 3 parts of Clostridium butyricum to the composite carrier, stir for 8 minutes, slowly spray the protective coloring solution, and continue stirring for 20 minutes. The temperature throughout the process is 33℃. S4. Feed the pellets into a twin-screw extruder, set the pelleting temperature to 80℃ and the screw speed to 220r / min, and use a 4.5mm diameter die and a 13mm cutter spacing to extrude and form pellets with a diameter of 4.5mm and a length of 13mm. S5, air-dry at 45℃ for 20 minutes, cool, sieve, and package to obtain a uniform, deep yellow composite microbial granule preparation.

[0040] Comparative Example The formula is made by adding a commercially available compound probiotic granule preparation. The strain composition, addition amount, and granulation temperature are completely consistent with those in Example 2. The only difference is that no modified starch carrier, compound high-temperature protection system, standardized granulation process, and coloring design are added.

[0041] Test case To verify the technical effect of the "high-temperature resistant composite microbial granule preparation" of the present invention, five experimental diets were set up.

[0042] The experimental group diet consisted of conventional commercial pig feed, supplemented with 0.3% of the high-temperature resistant compound microbial granule preparations prepared in Example 1 (Experimental Group 1), Example 2 (Experimental Group 2), and Example 3 (Experimental Group 3) of this invention. The blank control group diet (blank group) consisted of conventional commercial pig feed without any added probiotic preparations. Comparative diet (traditional control group): conventional commercial pig feed, with the addition of 0.3% of commercially available compound probiotic granules of the same strain (completely consistent with the strain composition, addition amount, and granulation temperature of Example 2 of this invention, the only difference being the absence of modified starch carrier, composite high-temperature protection system, standardized granulation process, and coloring design).

[0043] All experimental diets were uniformly processed using a high-temperature extrusion process at 70–80℃, with a screw speed of 210 r / min, and then air-dried at 42℃ after pelleting. Two hundred and fifty healthy, active three-way crossbred pigs (Duroc × Landrace × Large White) of similar weight and around 100 days old from a large-scale pig farm were randomly divided into five groups, with five replicates per group and ten pigs per replicate, corresponding to the five dietary treatments. All experimental pigs were fed in a standardized manner with free access to feed and water for a 60-day period.

[0044] The test was used to verify: (1) the heat resistance and activity retention of the strains in the formulation of the present invention under the high-temperature extrusion process at 70-80℃; (2) the conditioning effect of the formulation on the intestinal flora structure and intestinal health of pigs; (3) the optimization effect of the formulation on the growth performance and feed conversion ratio of pigs; and (4) the improvement effect of the formulation on the pig farming environment. The experimental data were analyzed using SPSS 26.0 statistical software with one-way ANOVA and Duncan's method for multiple comparisons. The experimental results are expressed as mean ± standard deviation.

[0045] (1) High-temperature resistance and activity of strains under high-temperature extrusion process at 70-80℃ The heat resistance activity of microbial strains under high-temperature extrusion processing at 70-80℃ is expressed as the viable cell survival rate. The viable cell survival rate refers to the percentage of viable functional bacteria that survive the entire pelleting process compared to the initial viable functional bacteria count in the pelleted feed. It directly reflects the ability of functional bacteria such as Bacillus coagulans, Bacillus licheniformis, and Clostridium butyricum to withstand the high-temperature and humid heat of pelleting. The calculation formula is as follows: Viable cell survival rate (%) = viable cell count after granulation ÷ viable cell count before granulation 100 The results of the viable bacteria survival rate determination are shown in Table 1.

[0046]

[0047] Table 1 Results of viable bacterial survival rate determination Note: Different letters under the same column indicate significant differences (P<0.05), while the same letter under the same column indicates no significant differences (P>0.05). The same applies to the following table; all data are based on air-dried data.

[0048] As shown in Table 1, after being extruded at a high temperature of 70-80℃, the survival rate of live bacteria in experimental groups 1, 2, and 3 of the present invention was significantly higher than that in the traditional control group (P < 0.05); there was no significant difference among the three experimental groups of the present invention (P > 0.05), indicating that the formulation parameters of the present invention are within the scope of protection of the claims, and the high temperature resistance is stable and universal, solving the defect of high temperature inactivation of traditional probiotics.

[0049] (2) Intestinal health indicators of pigs Intestinal health in pigs is represented by the daily average diarrhea rate, the rate of increase in lactic acid bacteria abundance, and the rate of decrease in Salmonella abundance.

[0050] Daily diarrhea rate (%) = Total number of diarrheal episodes in pigs / (Total number of pigs × Total number of days) × 100 Increase rate of lactic acid bacteria abundance in fecal samples (%) = (Absolute abundance of lactic acid bacteria in experimental group fecal samples - Absolute abundance of lactic acid bacteria in blank group fecal samples) / Absolute abundance of lactic acid bacteria in blank group fecal samples × 100 Fecal Salmonella abundance reduction rate (%) = (Absolute abundance of Salmonella in the control group - Absolute abundance of Salmonella in the experimental group) / Absolute abundance of Salmonella in the control group × 100 The results of the intestinal health index test in pigs are shown in Table 2.

[0051]

[0052] Table 2 Results of the intestinal health index test in pigs Table 2 shows that the average daily diarrhea rate of pigs in all experimental groups of this invention was significantly lower than that in the blank group and the traditional control group (P < 0.05), and the effects of increasing fecal lactic acid bacteria and inhibiting Salmonella were significantly better than those in the traditional control group (P < 0.05). Among them, experimental group 2 showed the best intestinal conditioning effect, which was consistent with the optimal compound formula. This proves that the three-strain compound system of this invention has significant inventiveness and synergistic effect.

[0053] (3) Pig production performance Pig production performance is expressed by average daily weight gain, average daily feed intake, and feed conversion ratio.

[0054] The results of the pig production performance test are shown in Table 3.

[0055]

[0056] Table 3 Results of pig production performance test Table 3 shows that there was no significant difference in average daily feed intake among the treatment groups (P > 0.05), thus excluding the interference variable of feed intake. The average daily weight gain of each experimental group of the present invention was significantly higher than that of the control group and the traditional control group (P < 0.05), and the feed conversion ratio was significantly lower than that of the blank group and the traditional control group (P < 0.05). This demonstrates that the formulation of the present invention can significantly improve feed utilization, reduce feed conversion ratio, and significantly increase average daily weight gain without increasing feed intake.

[0057] (4) Aquaculture environment indicators The environmental indicators for aquaculture are expressed by the decrease in ammonia nitrogen content and hydrogen sulfide concentration in excrement.

[0058] The results of the aquaculture environment index test are shown in Table 4.

[0059]

[0060] Table 4 Results of the Aquaculture Environmental Indicators Test As shown in Table 4, the ammonia nitrogen and hydrogen sulfide content in the excrement of each experimental group of the present invention were significantly lower than those of the blank group and the traditional control group (P<0.05), and the difference in the emission reduction effect of harmful gases was significant. It can effectively improve the pig farming environment and has outstanding advantages in green farming.

[0061] Based on the experimental results in Tables 1, 2, 3, and 4, the final conclusion can be drawn as follows: Compared to traditional probiotic formulations of the same strain, the high-temperature resistant composite microbial granule formulation of this invention has significant technical advantages in terms of high-temperature processing stability, intestinal health regulation, growth promotion and efficacy enhancement, and improvement of the breeding environment. Under a high-temperature extrusion granulation process of 70–80°C, the formulation of this invention can effectively retain high viable bacterial activity, with a significantly higher viable bacterial survival rate than traditional formulations, and the product performance is stable, overcoming the defect of traditional probiotics being easily inactivated at high temperatures.

[0062] This formulation can significantly reduce the diarrhea rate in pigs, optimize the intestinal flora structure, increase the abundance of beneficial bacteria, inhibit the growth of harmful bacteria, and has a prominent synergistic effect in intestinal conditioning. In livestock applications, it can significantly increase the average daily weight gain of pigs and reduce the feed conversion ratio without affecting their feed intake, effectively improving feed utilization and livestock production efficiency.

[0063] At the same time, the formulation can significantly reduce the content of harmful gases such as ammonia nitrogen and hydrogen sulfide in pig excrement, effectively improve the breeding environment, and has the dual advantages of stable production and efficiency and green environmental protection. The comprehensive application effect is significantly better than that of traditional similar products on the market.

[0064] The technical solutions of the present invention are not limited to the specific embodiments described above. Any technical modifications made in accordance with the technical solutions of the present invention fall within the protection scope of the present invention.

Claims

1. A high-temperature resistant composite microbial granule preparation, characterized in that: The product comprises the following raw materials by weight: 2-8 parts of live Bacillus coagulans powder, 1-5 parts of live Bacillus licheniformis powder, 0.6-3 parts of live Clostridium butyricum powder, 60-80 parts of modified corn starch, 10-20 parts of defatted rice bran, 2-5 parts of trehalose protectant, 0.2-0.8 parts of vitamin E stabilizer, 0.1-0.5 parts of natural plant colorant, and 5-10 parts of purified water.

2. The high-temperature resistant composite microbial granule preparation according to claim 1, characterized in that: The modified corn starch is hydroxypropyl modified corn starch, which has high temperature stability at 70-80℃ and a porous encapsulation structure; the trehalose and vitamin E constitute a composite high temperature protection system to achieve high activity retention of the strain under high temperature granulation; the high temperature resistant composite microbial granule preparation is prepared by twin-screw extrusion process, and its particle diameter is 2.0-4.5mm and particle length is 4-13mm.

3. A processing method for a high-temperature resistant composite microbial granule preparation, characterized in that: Includes the following steps: S1. Preparation of protective coloring solution: Under low temperature and sterile conditions, trehalose protective agent, vitamin E stabilizer and natural plant coloring agent are added to purified water in sequence, and stirred until the system is homogeneous and transparent. Let it stand for later use. S2. Preparation of composite porous carrier: Modified corn starch and defatted rice bran are mixed evenly in proportion to obtain composite porous carrier; S3. Inoculum embedding and mixing: Bacillus coagulans live powder, Bacillus licheniformis live powder and Clostridium butyricum live powder are evenly sprinkled into the composite porous carrier, protective coloring liquid is sprayed by atomization, and the inoculum is completely embedded by constant temperature stirring to obtain a moist mixture. S4. High-temperature extrusion granulation: The mixture is formed at high temperature through a twin-screw extrusion process to obtain pre-formed granules; S5. Low-temperature air drying and shaping: The initially shaped particles are air-dried at a constant temperature and cooled naturally to obtain composite microbial particles with stable properties. S6. Screening and Packaging: Screening removes unqualified particles, and selects uniform finished particles for storage in a light-proof and sealed manner.

4. The processing method of a high-temperature resistant composite microbial granule preparation according to claim 3, characterized in that: The mixing ratio of each component by weight is as follows: 2-8 parts of Bacillus coagulans live bacteria powder, 1-5 parts of Bacillus licheniformis live bacteria powder, 0.6-3 parts of Clostridium butyricum live bacteria powder, 60-80 parts of modified corn starch, 10-20 parts of defatted rice bran, 2-5 parts of trehalose protectant, 0.2-0.8 parts of vitamin E stabilizer, 0.1-0.5 parts of natural plant colorant, and 5-10 parts of purified water.

5. The processing method of a high-temperature resistant composite microbial granule preparation according to claim 3, characterized in that: In step S1, the preparation environment is a dust-free and sterile environment with an ambient temperature of 25-30℃, and the stirring time is 10-15 minutes. After the materials are completely dissolved, they are allowed to stand at low temperature for later use. In step S2, a horizontal mixer is used to stir at a uniform speed for 5 minutes to achieve a uniform mixture of modified corn starch and defatted rice bran.

6. The processing method of a high-temperature resistant composite microbial granule preparation according to claim 3, characterized in that: In step S3, the bacteria are first stirred for 8 minutes to achieve initial dispersion, and then the protective coloring liquid is sprayed by atomization. Stirring is continued for 15-20 minutes, and the temperature of the system is controlled at 33-35℃ throughout the process.

7. The processing method of a high-temperature resistant composite microbial granule preparation according to claim 3, characterized in that: In step S4, the granulation temperature is 70-80℃, the screw speed is 200-220r / min, and a die with a diameter of 2.0-4.5mm is used to extrude the pellets with a cutter spacing of 4-13mm.

8. The processing method of a high-temperature resistant composite microbial granule preparation according to claim 3, characterized in that: In step S5, the air-drying temperature is 40-45℃ and the air-drying time is 20-25 minutes. After removing the free moisture on the surface of the particles, the particles are allowed to cool naturally to room temperature.

9. The application of a high-temperature resistant composite microbial granule preparation, characterized in that, The compound microbial granule preparation described in any one of claims 1-2 can be directly added to pig compound feed for mixed feeding, which is suitable for large-scale breeding of pigs throughout the entire growth stage.

10. The application of the high-temperature resistant composite microbial granule preparation according to claim 9, characterized in that, The compound microbial granule preparation is added to the feed at a rate of 0.2%-0.5% of the total feed mass, and can be directly mixed and used without secondary processing.

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