Feed additive for degrading vomitoxin and application thereof

Abstract

The application relates to the technical field of feed additives, in particular to a feed additive for degrading vomitoxin and application, wherein the feed additive for degrading vomitoxin is composed of, in percentage by weight, Bacillus altitudinis G-1 bacterial liquid: 10-30%; solid carrier: 70-90%. Efficient and stress-resistant Bacillus altitudinis G-1 is obtained through screening, the fermentation liquor is mixed with the treated solid carrier, and low-temperature or spray drying is adopted for shaping. The strain can secrete specific degradation enzyme system to efficiently convert vomitoxin, and can maintain activity in processing and digestive tract environment by virtue of wide temperature range and wide pH tolerance characteristics; in combination with the protection and slow-release effect of the carrier, efficient degradation of vomitoxin and synchronous inhibition of mold growth are realized, and the problems of limited efficiency of existing degradation strains, unstable activity in complex environment and lack of mold regeneration inhibition capacity are solved.

Description

Technical Field

[0001] This application relates to the field of feed additive technology, and in particular to a feed additive that degrades vomitoxin and its application. Background Technology

[0002] Deoxynivalenol (DON), also known as deoxynivalenol, is a common mycotoxin produced by fungi of the genus Fusarium. It widely contaminates grains such as wheat, corn, and barley, as well as related feed products. DON is highly toxic and can cause severe damage to the digestive tract and immune system of animals, leading to symptoms such as vomiting, diarrhea, growth retardation, and decreased reproductive performance. It also poses a threat to human health through the food chain.

[0003] Currently, the main methods for controlling vomitoxin contamination include physical detoxification (such as adsorption, heating, and irradiation), chemical detoxification (such as acid-base treatment and oxidative degradation), and biological detoxification (such as microbial degradation and enzymatic hydrolysis). Physical detoxification methods often suffer from incomplete toxin removal and loss of nutrients; chemical detoxification methods may introduce harmful residues, affecting the safety of feed and food; while biological detoxification has advantages such as high specificity, mild conditions, and no secondary pollution, making it a current research hotspot.

[0004] Existing technologies for degrading vomitoxin mainly utilize Bacillus subtilis and lactic acid bacteria, but most strains suffer from limitations such as limited degradation efficiency, weak resistance, and poor stability in complex feed environments. For example, some strains can only achieve low-efficiency degradation under specific laboratory conditions and are difficult to adapt to the high temperatures, pH fluctuations, and harsh environments within the animal digestive tract during feed processing. *Bacillus glaber* (… Bacillus altitudinis As an important member of the Bacillus genus, it has the characteristics of strong stress resistance, rich metabolites and wide environmental adaptability. In particular, it has significant advantages in enzyme production capacity and tolerance to extreme environments. However, this potential has not yet been fully applied to the field of vomitoxin detoxification.

[0005] Therefore, screening out a strain of Bacillus altissima that efficiently degrades vomitoxin and developing its application technology is of great practical significance for solving the problem of feed toxin contamination and ensuring feed safety. Summary of the Invention

[0006] This application provides a feed additive for degrading vomitoxin and its application, in order to solve the problems of limited efficiency of existing degrading strains, unstable activity in complex environments, and lack of ability to inhibit mold regeneration.

[0007] This application provides a feed additive for degrading vomitoxin, which, by weight percentage, comprises: Highland Bacillus G-1 bacterial suspension: 10-30%; Solid carrier: 70-90%.

[0008] Optionally, the effective viable count of the *Bacillus hygroscopicus* strain G-1 in the bacterial suspension is not less than 5.0 × 10⁻⁶. 9 CFU / mL.

[0009] Optionally, the Bacillus hygroscopicus strain G-1 is capable of growing and degrading vomitoxin under conditions of 27-67°C and pH 5.0-9.0.

[0010] Optionally, the solid carrier is one or more of wheat bran, corn flour, rice husk powder, zeolite powder, montmorillonite, or diatomaceous earth.

[0011] Optionally, the method for screening and obtaining the Bacillus hygroscopicus strain G-1 includes the following steps: S1. Separation and purification: Take 10 g of the sample containing microorganisms and place it in 100 mL of sterile water. Shake at 37℃ and 180 r / min for 1 h. Take the supernatant and perform a 10-fold serial dilution. Select 10 g of the supernatant for further dilution. -4 Up to 10 -7 Diluted bacterial suspensions were spread on NA plates and incubated at 37°C for 24 h. Single colonies were picked and subjected to at least three streak purification cultures to obtain pure cultures. S2. Initial screening of degradation performance: The pure culture was inoculated into NB medium and cultured at 37℃ and 180 r / min for 24 h. 950 μL of fermentation supernatant was mixed with 50 μL of vomitoxin standard solution with a concentration of 100 μg / mL and co-cultured at 60℃ for 72 h. The absorbance was measured at wavelengths of 450 / 630 nm using an enzyme-linked immunosorbent assay kit. The residual amount of vomitoxin and the degradation rate were calculated, and strains with high degradation rates were screened. S3. Strain identification: Genomic DNA was extracted from the strain, and PCR amplification was performed using universal primers 27F and 1492R for the bacterial 16S rRNA gene. The product was sequenced, and the sequence was compared with the NCBI database using BLAST. Combined with physiological and biochemical characteristics, the target strain G-1 with 100% homology to Bacillus hygroscopicus was obtained.

[0012] This application also proposes a method for preparing a feed additive that degrades vomitoxin, comprising the following steps: S1. Seed culture preparation: Bacillus hygroscopicus strain G-1 was inoculated into a shake flask containing NB medium and cultured with shaking at 37℃ and 180 rpm for 14-24 h to obtain seed culture; S2. Fermentation Culture: The seed culture is inoculated into a fermenter containing fermentation medium at a volume inoculation rate of 5-10%, and fermented at 37℃, 180 rpm, and an aeration rate of 0.5-1.0 vvm for 24-48 h to obtain the fermentation broth. The effective viable cell count of the fermentation broth is not less than 5.0 × 10⁻⁶. 9 CFU / mL.

[0013] S3. Carrier pretreatment: The solid carrier is crushed by a pulverizer and passed through an 80-mesh sieve. The material on the sieve is returned to be crushed again and then sterilized by moist heat at 105-121℃ for 15-30 min. After cooling to room temperature, the pretreated solid carrier is obtained. S4. Mixing and post-processing: The fermentation broth and the pretreated solid carrier are mixed at a weight ratio of 10-30%:70-90%, and stirred at 30-50 rpm for 20-40 min using a twin-screw mixer until the coefficient of variation of the mixing uniformity is ≤10%. The uniformly mixed material is then dried at 40-50℃ until the moisture content is ≤12% to obtain a powdered feed additive.

[0014] Optionally, in step S1, the NB culture medium has the following composition: 10 g / L peptone, 3 g / L beef extract, 5 g / L sodium chloride, water as solvent, and a pH value of 7.0-7.2.

[0015] Optionally, in step S3, the solid carrier is bran, and its pretreatment includes crushing to a particle size ≤0.5 mm and adjusting the moisture content to 10-15%.

[0016] Optionally, in step S4, the drying is performed at a low temperature of 40-50°C or by spray drying.

[0017] This application also proposes the application of a feed additive that degrades vomitoxin, used to reduce the vomitoxin content in grains or feed and inhibit mold growth.

[0018] Therefore, this application has at least the following beneficial effects: The *Bacillus hygroscopicus* G-1 strain obtained in this application can secrete an enzyme system that degrades vomitoxin. Under suitable conditions, the degradation rate of vomitoxin exceeds 85% within 48 hours, converting vomitoxin into low-toxicity or non-toxic metabolites. Simultaneously, the bacterial cell's own secondary metabolic system can further convert these products and incorporate them into the carbon metabolic stream, thereby achieving a complete transformation from toxin decomposition to complete assimilation, eliminating the risk of intermediate toxic substance accumulation and toxin regeneration. The strain G-1 used in this application exhibits excellent environmental adaptability, a characteristic stemming from the adjustable composition of its cell membrane lipids and the efficient expression of protective mechanisms such as heat shock proteins and acid-base tolerance enzymes. This strain maintains metabolic activity across a wide temperature range of 27°C to 67°C. The key lies in its ability to maintain conformational stability of intracellular enzyme proteins even under high temperatures, allowing it to tolerate short-term high-temperature treatments of 80°C to 90°C during feed pelleting. Simultaneously, the strain possesses a strong ability to regulate intracellular pH homeostasis, effectively performing degradation functions within a pH range of 5.0 (simulating gastric acid) to 9.0 (simulating intestinal alkaline environment), thus ensuring its survival and continued function throughout the entire animal digestive tract chain, including feed processing, storage, and more. The feed additive provided in this application not only efficiently degrades existing vomitoxin, but also competitively utilizes oxygen, moisture, and nutrients in the feed through the rapid colonization and proliferation of strain G-1, thereby inhibiting the growth space of molds from an ecological niche perspective. Furthermore, the bacteriocins, organic acids (such as lactic acid and acetic acid), and volatile antibacterial substances produced by the strain's metabolism can disrupt the cell membrane structure of molds, inhibiting spore germination and hyphal extension. This dual mechanism of "nutrient competition" and "metabolic inhibition" can effectively curb the reproduction of toxin-producing molds such as Fusarium from the source, blocking the continuous synthesis and accumulation of vomitoxin. This application's embodiments optimize the pretreatment process of solid carriers (such as wheat bran), precisely controlling their moisture content and particle size distribution, and sterilizing them to construct a microenvironment conducive to bacterial adsorption and colonization, while providing physical protection. The carrier not only serves as a physical support for the bacteria, but its porous structure also adsorbs metabolic products and moisture, providing stable protection for the bacteria (especially spore morphology) during storage. In practical applications, the carrier gradually decomposes in feed or fermentation systems, achieving slow release and uniform dispersion of the bacteria, thereby extending its action time, expanding its range of action, and significantly improving the convenience and sustainability of the product's effects. This application integrates highly efficient natural screening strains, agricultural by-product carriers, and a mild fermentation process to construct a complete bio-based detoxification solution. This system does not rely on chemical adsorbents, does not introduce exogenous harmful residues, and avoids nutrient loss or secondary pollution that may occur with traditional physicochemical methods. Strain G-1 and its metabolites are safe for animals, and some metabolites, such as organic acids, even have auxiliary functions in improving intestinal health. This solution closely aligns with the actual conditions of the feed industry and livestock production, with readily available raw materials, a simple process, and strong compatibility. While ensuring fundamental feed safety, it achieves environmental friendliness and cost control, demonstrating significant prospects for industrialization and promotion.

[0019] This addresses the problems of existing degradation strains having limited efficiency, unstable activity in complex environments, and a lack of ability to inhibit mold regeneration.

[0020] Additional aspects and advantages of this application will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of this application. Attached Figure Description

[0021] The above and / or additional aspects and advantages of this application will become apparent and readily understood from the following description of the embodiments taken in conjunction with the accompanying drawings, wherein: Figure 1 This is a flowchart illustrating a method for preparing a feed additive that degrades vomitoxin, according to an embodiment of this application. Figure 2 The images show the colony and cell morphology of strain G-1 provided in the embodiments of this application. Figure 3 This is a phylogenetic tree of strain G-1 provided according to the embodiments of this application; Figure 4 This is an example diagram illustrating the effect of temperature on the growth of strain G-1 according to embodiments of this application; Figure 5 This is an example diagram illustrating the effect of pH on the growth of strain G-1 according to embodiments of this application; Figure 6 This is an example diagram illustrating the effect of strain G-1 provided in the embodiments of this application on the vomitoxin content in fermented wheat bran; Figure 7 This is an example diagram illustrating the changes in pH, bacteria, mold, and lactic acid bacteria counts of bran during fermentation according to embodiments of this application. Detailed Implementation

[0022] The technical solutions in the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of this application without creative effort are within the scope of protection of this application.

[0023] In the embodiments of this application, unless otherwise specified, the raw materials or processing techniques are conventional commercially available raw materials or conventional processing techniques in the art.

[0024] The present application will now be described with reference to specific embodiments. It should be noted that these embodiments are merely descriptive and do not limit the present application in any way.

[0025] In this embodiment of the application, the drug source includes: Bacillus hygroscopicus, specifically Bacillus hygroscopicus G-1, which is deposited at the China General Microbiological Culture Collection Center with accession number CGMCC NO.37321.

[0026] Example 1 This application provides a feed additive for degrading vomitoxin, which, by weight percentage, comprises: Highland Bacillus G-1 bacterial suspension: 10%; Solid carrier: 90%.

[0027] Among them, the effective viable count of Bacillus hygroscopicus strain G-1 in bacterial suspension is not less than 5.0 × 10⁻⁶. 9 CFU / mL.

[0028] Among them, Bacillus hygroscopicus strain G-1 can grow and degrade vomitoxin under conditions of 27-67℃ and pH 5.0-9.0.

[0029] The solid carrier is wheat bran.

[0030] The method for screening and obtaining Bacillus subtilis strain G-1 includes the following steps: S1. Separation and purification: Take 10 g of the sample containing microorganisms and place it in 100 mL of sterile water. Shake at 37℃ and 180 r / min for 1 h. Take the supernatant and perform a 10-fold serial dilution. Select 10 g of the supernatant for further dilution. -4 Up to 10 -7 Diluted bacterial suspensions were spread on NA plates and incubated at 37°C for 24 h. Single colonies were picked and subjected to at least three streak purification cultures to obtain pure cultures. S2. Initial screening of degradation performance: The pure culture was inoculated into NB medium and cultured at 37℃ and 180 r / min for 24 h. 950 μL of fermentation supernatant was mixed with 50 μL of vomitoxin standard solution with a concentration of 100 μg / mL and co-cultured at 60℃ for 72 h. The absorbance was measured at wavelengths of 450 / 630 nm using an enzyme-linked immunosorbent assay kit. The residual amount of vomitoxin and the degradation rate were calculated, and strains with high degradation rates were screened. S3. Strain identification: Genomic DNA was extracted from the strain, and PCR amplification was performed using universal primers 27F and 1492R for the bacterial 16S rRNA gene. The product was sequenced, and the sequence was compared with the NCBI database using BLAST. Combined with physiological and biochemical characteristics, the target strain G-1 with 100% homology to Bacillus hygroscopicus was obtained.

[0031] This application also proposes a method for preparing a feed additive that degrades vomitoxin, such as... Figure 1 As shown, it includes the following steps: S1. Seed culture preparation: Bacillus hygroscopicus strain G-1 was inoculated into a shake flask containing NB medium and cultured with shaking at 37℃ and 180 rpm for 14 h to obtain seed culture; Understandably, the shake-flask seed culture in this embodiment aims to achieve the initial activation and proliferation of strain G-1 from its storage state to a highly active growth state. Strict control of the culture temperature and shaking rate provides optimal growth kinetics for the cells, ensuring a metabolically vigorous and highly pure seed culture. This step is fundamental to the successful large-scale fermentation subsequent; a high-quality seed culture significantly shortens the fermentation lag period, ensuring the fermentation broth ultimately reaches the preset high viable cell count, providing a source guarantee for the preparation of highly efficient detoxified products.

[0032] S2. Fermentation Culture: Inoculate the seed culture at a volume of 5% into a fermenter containing fermentation medium. Ferment and culture at 37℃, 180 rpm, and an aeration rate of 0.5 vvm for 24 h to obtain the fermentation broth. The effective viable cell count of the fermentation broth should not be less than 5.0 × 10⁻⁶. 9 CFU / mL; Understandably, this application embodiment uses a fermenter for large-scale cultivation. By precisely controlling the inoculum size, temperature, stirring rate, and aeration rate, an optimized environment for the large-scale, high-density growth of *Bacillus hygroscopicus* G-1 is created. An appropriate inoculum size facilitates the rapid establishment of growth dominance, while adequate aeration and stirring ensure a uniform supply of dissolved oxygen and nutrients, thereby maximizing the accumulation of bacterial biomass and metabolites. This step is crucial for obtaining a high-concentration, high-activity fermentation broth, directly determining the effective dosage and degradation potential of the functional strains in the final additive.

[0033] S3. Carrier pretreatment: The solid carrier is crushed by a pulverizer and passed through an 80-mesh sieve. The material on the sieve is returned to be crushed again. The moisture content is adjusted to 10%, and the carrier is sterilized by moist heat at 105℃ for 15 min. It is then cooled to room temperature for use to obtain the pretreated solid carrier. It is understood that the pretreatment of the solid carrier in this embodiment, including pulverization, sieving, water adjustment, and sterilization, aims to create a suitable microenvironment for bacterial adsorption and survival. Controlling the carrier particle size and water content optimizes its physical structure, improving its compatibility and carrying capacity with the liquid fermentation broth. Strict moist heat sterilization completely eliminates any contaminating bacteria carried by the carrier itself, preventing them from contaminating the product and competing with functional strains. This step ensures the purity and applicability of the carrier as a bacterial "warehouse," and is a crucial pretreatment for preparing stable and homogeneous solid additives.

[0034] S4. Mixing and post-processing: The fermentation broth and the pretreated solid carrier are mixed at a weight ratio of 10%:90%, and stirred at 30 rpm for 20 min using a twin-screw mixer until the coefficient of variation of the mixing uniformity is ≤10%. The uniformly mixed material is then dried at 40℃ until the moisture content is ≤12% to obtain a powdered feed additive.

[0035] It is understood that the embodiments of this application, by controlling the mixing ratio of bacterial solution and carrier, employing mechanical mixing at specific speeds and times, and finally performing gentle drying, aim to achieve efficient, uniform loading and stable solidification of functional strains on the carrier. Precise mixing ratios are fundamental to balancing product efficacy and cost; gentle yet thorough mixing ensures uniform bacterial dispersion; the subsequent low-temperature or rapid drying process aims to remove excess moisture for preservation while minimizing thermal damage to bacterial activity. This step ultimately determines the product's physical properties, viable cell survival rate, and storage stability, and is the final step in obtaining a high-quality, directly applicable powdered feed additive.

[0036] In step S1, the NB culture medium consists of: 10 g / L peptone, 3 g / L beef extract, 5 g / L sodium chloride, water as solvent, and a pH of 7.0.

[0037] In step S3, the solid carrier is bran, and its pretreatment includes crushing to a particle size ≤0.5 mm and adjusting the moisture content to 10%.

[0038] In step S4, drying is performed at a low temperature of 40°C.

[0039] This application also proposes the application of a feed additive that degrades vomitoxin, used to reduce the vomitoxin content in grains or feed and inhibit mold growth.

[0040] Example 2 This application provides a feed additive for degrading vomitoxin, which, by weight percentage, comprises: Highland Bacillus G-1 bacterial suspension: 15%; Solid carrier: 85%.

[0041] Among them, the effective viable count of Bacillus hygroscopicus strain G-1 in bacterial suspension is not less than 5.0 × 10⁻⁶. 9 CFU / mL.

[0042] Among them, Bacillus hygroscopicus strain G-1 can grow and degrade vomitoxin under conditions of 27-67℃ and pH 5.0-9.0.

[0043] The solid carrier is wheat bran.

[0044] The method for screening and obtaining Bacillus subtilis strain G-1 includes the following steps: S1. Separation and purification: Take 10 g of the sample containing microorganisms and place it in 100 mL of sterile water. Shake at 37℃ and 180 r / min for 1 h. Take the supernatant and perform a 10-fold serial dilution. Select 10 g of the supernatant for further dilution. -4 Up to 10 -7Diluted bacterial suspensions were spread on NA plates and incubated at 37°C for 24 hours. Single colonies were picked and subjected to at least three streak purification cultures to obtain pure cultures. S2. Initial screening of degradation performance: The pure culture was inoculated into NB medium and cultured at 37℃ and 180 r / min for 24 h. 950 μL of fermentation supernatant was mixed with 50 μL of vomitoxin standard solution with a concentration of 100 μg / mL and co-cultured at 60℃ for 72 h. The absorbance was measured at wavelengths of 450 / 630 nm using an enzyme-linked immunosorbent assay kit. The residual amount of vomitoxin and the degradation rate were calculated, and strains with high degradation rates were screened. S3. Strain identification: Genomic DNA was extracted from the strain, and PCR amplification was performed using universal primers 27F and 1492R for the bacterial 16S rRNA gene. The product was sequenced, and the sequence was compared with the NCBI database using BLAST. Combined with physiological and biochemical characteristics, the target strain G-1 with 100% homology to Bacillus hygroscopicus was obtained.

[0045] This application also proposes a method for preparing a feed additive that degrades vomitoxin, such as... Figure 1 As shown, it includes the following steps: S1. Seed culture preparation: Bacillus hygroscopicus strain G-1 was inoculated into a shake flask containing NB medium and cultured with shaking at 37℃ and 180 rpm for 18 h to obtain seed culture; Understandably, the shake-flask seed culture in this embodiment aims to achieve the initial activation and proliferation of strain G-1 from its storage state to a highly active growth state. Strict control of the culture temperature and shaking rate provides optimal growth kinetics for the cells, ensuring a metabolically vigorous and highly pure seed culture. This step is fundamental to the successful large-scale fermentation subsequent; a high-quality seed culture significantly shortens the fermentation lag period, ensuring the fermentation broth ultimately reaches the preset high viable cell count, providing a source guarantee for the preparation of highly efficient detoxified products.

[0046] S2. Fermentation Culture: The seed culture was inoculated into a fermenter containing fermentation medium at a volume inoculation rate of 7.5%. Fermentation was carried out at 37℃, 180 rpm, and an aeration rate of 0.75 vvm for 36 h to obtain the fermentation broth. The effective viable cell count of the fermentation broth was not less than 5.0 × 10⁻⁶. 9 CFU / mL; Understandably, this application embodiment uses a fermenter for large-scale cultivation. By precisely controlling the inoculum size, temperature, stirring rate, and aeration rate, an optimized environment for the large-scale, high-density growth of *Bacillus hygroscopicus* G-1 is created. An appropriate inoculum size facilitates the rapid establishment of growth dominance, while adequate aeration and stirring ensure a uniform supply of dissolved oxygen and nutrients, thereby maximizing the accumulation of bacterial biomass and metabolites. This step is crucial for obtaining a high-concentration, high-activity fermentation broth, directly determining the effective dosage and degradation potential of the functional strains in the final additive.

[0047] S3. Carrier pretreatment: The solid carrier is crushed by a pulverizer and then passed through an 80-mesh sieve. The material on the sieve is returned to be crushed again. The moisture content is adjusted to 12.5%, and the carrier is sterilized by moist heat at 113℃ for 22.5 min. After cooling to room temperature, the pretreated solid carrier is obtained. It is understood that the pretreatment of the solid carrier in this embodiment, including pulverization, sieving, water adjustment, and sterilization, aims to create a suitable microenvironment for bacterial adsorption and survival. Controlling the carrier particle size and water content optimizes its physical structure, improving its compatibility and carrying capacity with the liquid fermentation broth. Strict moist heat sterilization completely eliminates any contaminating bacteria carried by the carrier itself, preventing them from contaminating the product and competing with functional strains. This step ensures the purity and applicability of the carrier as a bacterial "warehouse," and is a crucial pretreatment for preparing stable and homogeneous solid additives.

[0048] S4. Mixing and post-processing: The fermentation broth and the pretreated solid carrier are mixed at a weight ratio of 15%:85%, and stirred at 40 rpm for 30 min using a twin-screw mixer until the coefficient of variation of the mixing uniformity is ≤10%. The uniformly mixed material is then dried at 45℃ until the moisture content is ≤12% to obtain a powdered feed additive.

[0049] It is understood that the embodiments of this application, by controlling the mixing ratio of bacterial solution and carrier, employing mechanical mixing at specific speeds and times, and finally performing gentle drying, aim to achieve efficient, uniform loading and stable solidification of functional strains on the carrier. Precise mixing ratios are fundamental to balancing product efficacy and cost; gentle yet thorough mixing ensures uniform bacterial dispersion; the subsequent low-temperature or rapid drying process aims to remove excess moisture for preservation while minimizing thermal damage to bacterial activity. This step ultimately determines the product's physical properties, viable cell survival rate, and storage stability, and is the final step in obtaining a high-quality, directly applicable powdered feed additive.

[0050] In step S1, the NB culture medium consists of: 10 g / L peptone, 3 g / L beef extract, 5 g / L sodium chloride, water as solvent, and a pH of 7.0.

[0051] In step S3, the solid carrier is bran, and its pretreatment includes crushing to a particle size ≤0.5 mm and adjusting the moisture content to 12.5%.

[0052] In step S4, drying is performed at a low temperature of 45°C.

[0053] This application also proposes the application of a feed additive that degrades vomitoxin, used to reduce the vomitoxin content in grains or feed and inhibit mold growth.

[0054] Example 3 This application provides a feed additive for degrading vomitoxin, which, by weight percentage, comprises: Highland Bacillus G-1 bacterial suspension: 20%; Solid carrier: 80%.

[0055] Among them, the effective viable count of Bacillus hygroscopicus strain G-1 in bacterial suspension is not less than 5.0 × 10⁻⁶. 9 CFU / mL.

[0056] Among them, Bacillus hygroscopicus strain G-1 can grow and degrade vomitoxin under conditions of 27-67℃ and pH 5.0-9.0.

[0057] The solid carrier is wheat bran.

[0058] The method for screening and obtaining Bacillus subtilis strain G-1 includes the following steps: S1. Separation and purification: Take 10 g of the sample containing microorganisms and place it in 100 mL of sterile water. Shake at 37℃ and 180 r / min for 1 h. Take the supernatant and perform a 10-fold serial dilution. Select 10 g of the supernatant for further dilution. -4 Up to 10 -7 Diluted bacterial suspensions were spread on NA plates and incubated at 37°C for 24 h. Single colonies were picked and subjected to at least three streak purification cultures to obtain pure cultures. S2. Initial screening of degradation performance: The pure culture was inoculated into NB medium and cultured at 37℃ and 180 r / min for 24 h. 950 μL of fermentation supernatant was mixed with 50 μL of vomitoxin standard solution with a concentration of 100 μg / mL and co-cultured at 60℃ for 72 h. The absorbance was measured at wavelengths of 450 / 630 nm using an enzyme-linked immunosorbent assay kit. The residual amount of vomitoxin and the degradation rate were calculated, and strains with high degradation rates were screened. S3. Strain identification: Genomic DNA was extracted from the strain, and PCR amplification was performed using universal primers 27F and 1492R for the bacterial 16S rRNA gene. The product was sequenced, and the sequence was compared with the NCBI database using BLAST. Combined with physiological and biochemical characteristics, the target strain G-1 with 100% homology to Bacillus hygroscopicus was obtained.

[0059] This application also proposes a method for preparing a feed additive that degrades vomitoxin, such as... Figure 1 As shown, it includes the following steps: S1. Seed culture preparation: Bacillus hygroscopicus strain G-1 was inoculated into a shake flask containing NB medium and cultured with shaking at 37℃ and 180 rpm for 22 h to obtain seed culture; Understandably, the shake-flask seed culture in this embodiment aims to achieve the initial activation and proliferation of strain G-1 from its storage state to a highly active growth state. Strict control of the culture temperature and shaking rate provides optimal growth kinetics for the cells, ensuring a metabolically vigorous and highly pure seed culture. This step is fundamental to the successful large-scale fermentation subsequent; a high-quality seed culture significantly shortens the fermentation lag period, ensuring the fermentation broth ultimately reaches the preset high viable cell count, providing a source guarantee for the preparation of highly efficient detoxified products.

[0060] S2. Fermentation Culture: The seed culture was inoculated into a fermenter containing fermentation medium at a volume of 10%. Fermentation was carried out at 37℃, 180 rpm, and an aeration rate of 1.0 vvm for 48 h to obtain the fermentation broth. The effective viable cell count of the fermentation broth was not less than 5.0 × 10⁻⁶. 9 CFU / mL; Understandably, this application embodiment uses a fermenter for large-scale cultivation. By precisely controlling the inoculum size, temperature, stirring rate, and aeration rate, an optimized environment for the large-scale, high-density growth of *Bacillus hygroscopicus* G-1 is created. An appropriate inoculum size facilitates the rapid establishment of growth dominance, while adequate aeration and stirring ensure a uniform supply of dissolved oxygen and nutrients, thereby maximizing the accumulation of bacterial biomass and metabolites. This step is crucial for obtaining a high-concentration, high-activity fermentation broth, directly determining the effective dosage and degradation potential of the functional strains in the final additive.

[0061] S3. Carrier pretreatment: The solid carrier is crushed by a pulverizer and passed through an 80-mesh sieve. The material on the sieve is returned to be crushed again. The moisture content is adjusted to 15%, and the carrier is sterilized by moist heat at 121℃ for 30 min. It is then cooled to room temperature for use to obtain the pretreated solid carrier. It is understood that the pretreatment of the solid carrier in this embodiment, including pulverization, sieving, water adjustment, and sterilization, aims to create a suitable microenvironment for bacterial adsorption and survival. Controlling the carrier particle size and water content optimizes its physical structure, improving its compatibility and carrying capacity with the liquid fermentation broth. Strict moist heat sterilization completely eliminates any contaminating bacteria carried by the carrier itself, preventing them from contaminating the product and competing with functional strains. This step ensures the purity and applicability of the carrier as a bacterial "warehouse," and is a crucial pretreatment for preparing stable and homogeneous solid additives.

[0062] S4. Mixing and post-processing: The fermentation broth and the pretreated solid carrier are mixed at a weight ratio of 20%:80%, and stirred at 50 rpm for 40 min using a twin-screw mixer until the coefficient of variation of the mixing uniformity is ≤10%. The uniformly mixed material is then spray-dried at 50℃ until the moisture content is ≤12% to obtain a powdered feed additive.

[0063] It is understood that the embodiments of this application, by controlling the mixing ratio of bacterial solution and carrier, employing mechanical mixing at specific speeds and times, and finally performing gentle drying, aim to achieve efficient, uniform loading and stable solidification of functional strains on the carrier. Precise mixing ratios are fundamental to balancing product efficacy and cost; gentle yet thorough mixing ensures uniform bacterial dispersion; the subsequent low-temperature or rapid drying process aims to remove excess moisture for preservation while minimizing thermal damage to bacterial activity. This step ultimately determines the product's physical properties, viable cell survival rate, and storage stability, and is the final step in obtaining a high-quality, directly applicable powdered feed additive.

[0064] In step S1, the NB culture medium consists of: 10 g / L peptone, 3 g / L beef extract, 5 g / L sodium chloride, water as solvent, and a pH of 7.0.

[0065] In step S3, the solid carrier is wheat bran, and its pretreatment includes crushing to a particle size ≤0.5mm and adjusting the moisture content to 15%.

[0066] In step S4, the drying process is spray drying.

[0067] This application also proposes the application of a feed additive that degrades vomitoxin, used to reduce the vomitoxin content in grains or feed and inhibit mold growth.

[0068] Example 4 This application provides a feed additive for degrading vomitoxin, which, by weight percentage, comprises: Highland Bacillus G-1 bacterial culture: 25%; Solid carrier: 75%.

[0069] Among them, the effective viable count of Bacillus hygroscopicus strain G-1 in bacterial suspension is not less than 5.0 × 10⁻⁶. 9 CFU / mL.

[0070] Among them, Bacillus hygroscopicus strain G-1 can grow and degrade vomitoxin under conditions of 27-67℃ and pH 5.0-9.0.

[0071] The solid carrier is a mixture of corn flour and zeolite powder in a mass ratio of 1:1.

[0072] The method for screening and obtaining Bacillus subtilis strain G-1 includes the following steps: S1. Separation and purification: Take 10 g of the sample containing microorganisms and place it in 100 mL of sterile water. Shake at 37℃ and 180 r / min for 1 h. Take the supernatant and perform a 10-fold serial dilution. Select 10 g of the supernatant for further dilution. -4 Up to 10 -7 Diluted bacterial suspensions were spread on NA plates and incubated at 37°C for 24 h. Single colonies were picked and subjected to at least three streak purification cultures to obtain pure cultures. S2. Initial screening of degradation performance: The pure culture was inoculated into NB medium and cultured at 37℃ and 180 r / min for 24 h. 950 μL of fermentation supernatant was mixed with 50 μL of vomitoxin standard solution with a concentration of 100 μg / mL and co-cultured at 60℃ for 72 h. The absorbance was measured at wavelengths of 450 / 630 nm using an enzyme-linked immunosorbent assay kit. The residual amount of vomitoxin and the degradation rate were calculated, and strains with high degradation rates were screened. S3. Strain identification: Genomic DNA was extracted from the strain, and PCR amplification was performed using universal primers 27F and 1492R for the bacterial 16S rRNA gene. The product was sequenced, and the sequence was compared with the NCBI database using BLAST. Combined with physiological and biochemical characteristics, the target strain G-1 with 100% homology to Bacillus hygroscopicus was obtained.

[0073] This application also proposes a method for preparing a feed additive that degrades vomitoxin, such as... Figure 1 As shown, it includes the following steps: S1. Seed culture preparation: Bacillus hygroscopicus strain G-1 was inoculated into a shake flask containing NB medium and cultured with shaking at 37℃ and 180 rpm for 14 h to obtain seed culture; Understandably, the shake-flask seed culture in this embodiment aims to achieve the initial activation and proliferation of strain G-1 from its storage state to a highly active growth state. Strict control of the culture temperature and shaking rate provides optimal growth kinetics for the cells, ensuring a metabolically vigorous and highly pure seed culture. This step is fundamental to the successful large-scale fermentation subsequent; a high-quality seed culture significantly shortens the fermentation lag period, ensuring the fermentation broth ultimately reaches the preset high viable cell count, providing a source guarantee for the preparation of highly efficient detoxified products.

[0074] S2. Fermentation Culture: Inoculate the seed culture at a volume of 5% into a fermenter containing fermentation medium. Ferment and culture at 37℃, 180 rpm, and an aeration rate of 0.5 vvm for 24 h to obtain the fermentation broth. The effective viable cell count of the fermentation broth should not be less than 5.0 × 10⁻⁶. 9 CFU / mL; Understandably, this application embodiment uses a fermenter for large-scale cultivation. By precisely controlling the inoculum size, temperature, stirring rate, and aeration rate, an optimized environment for the large-scale, high-density growth of *Bacillus hygroscopicus* G-1 is created. An appropriate inoculum size facilitates the rapid establishment of growth dominance, while adequate aeration and stirring ensure a uniform supply of dissolved oxygen and nutrients, thereby maximizing the accumulation of bacterial biomass and metabolites. This step is crucial for obtaining a high-concentration, high-activity fermentation broth, directly determining the effective dosage and degradation potential of the functional strains in the final additive.

[0075] S3. Carrier pretreatment: The solid carrier is crushed by a pulverizer and passed through an 80-mesh sieve. The material on the sieve is returned to be crushed again. The moisture content is adjusted to 10%, and the carrier is sterilized by moist heat at 105℃ for 15 min. It is then cooled to room temperature for use to obtain the pretreated solid carrier. It is understood that the pretreatment of the solid carrier in this embodiment, including pulverization, sieving, water adjustment, and sterilization, aims to create a suitable microenvironment for bacterial adsorption and survival. Controlling the carrier particle size and water content optimizes its physical structure, improving its compatibility and carrying capacity with the liquid fermentation broth. Strict moist heat sterilization completely eliminates any contaminating bacteria carried by the carrier itself, preventing them from contaminating the product and competing with functional strains. This step ensures the purity and applicability of the carrier as a bacterial "warehouse," and is a crucial pretreatment for preparing stable and homogeneous solid additives.

[0076] S4. Mixing and post-processing: The fermentation broth and the pretreated solid carrier are mixed at a weight ratio of 25%:75%, and stirred at 30 rpm for 20 min using a twin-screw mixer until the coefficient of variation of the mixing uniformity is ≤10%. The uniformly mixed material is then dried at 40℃ until the moisture content is ≤12% to obtain a powdered feed additive.

[0077] It is understood that the embodiments of this application, by controlling the mixing ratio of bacterial solution and carrier, employing mechanical mixing at specific speeds and times, and finally performing gentle drying, aim to achieve efficient, uniform loading and stable solidification of functional strains on the carrier. Precise mixing ratios are fundamental to balancing product efficacy and cost; gentle yet thorough mixing ensures uniform bacterial dispersion; the subsequent low-temperature or rapid drying process aims to remove excess moisture for preservation while minimizing thermal damage to bacterial activity. This step ultimately determines the product's physical properties, viable cell survival rate, and storage stability, and is the final step in obtaining a high-quality, directly applicable powdered feed additive.

[0078] In step S1, the NB culture medium consists of: 10 g / L peptone, 3 g / L beef extract, 5 g / L sodium chloride, water as solvent, and a pH of 7.0.

[0079] In step S4, drying is performed at a low temperature of 40°C.

[0080] This application also proposes the application of a feed additive that degrades vomitoxin, used to reduce the vomitoxin content in grains or feed and inhibit mold growth.

[0081] Example 5 This application provides a feed additive for degrading vomitoxin, which, by weight percentage, comprises: Highland Bacillus G-1 bacterial suspension: 30%; Solid carrier: 70%.

[0082] Among them, the effective viable count of Bacillus hygroscopicus strain G-1 in bacterial suspension is not less than 5.0 × 10⁻⁶. 9 CFU / mL.

[0083] Among them, Bacillus hygroscopicus strain G-1 can grow and degrade vomitoxin under conditions of 27-67℃ and pH 5.0-9.0.

[0084] The solid carrier is a mixture of rice husk powder and montmorillonite in a mass ratio of 2:1.

[0085] The method for screening and obtaining Bacillus subtilis strain G-1 includes the following steps: S1. Separation and purification: Take 10 g of the sample containing microorganisms and place it in 100 mL of sterile water. Shake at 37℃ and 180 r / min for 1 h. Take the supernatant and perform a 10-fold serial dilution. Select 10 g of the supernatant for further dilution. -4 Up to 10 -7 Diluted bacterial suspensions were spread on NA plates and incubated at 37°C for 24 h. Single colonies were picked and subjected to at least three streak purification cultures to obtain pure cultures. S2. Initial screening of degradation performance: The pure culture was inoculated into NB medium and cultured at 37℃ and 180 r / min for 24 h. 950 μL of fermentation supernatant was mixed with 50 μL of vomitoxin standard solution with a concentration of 100 μg / mL and co-cultured at 60℃ for 72 h. The absorbance was measured at wavelengths of 450 / 630 nm using an enzyme-linked immunosorbent assay kit. The residual amount of vomitoxin and the degradation rate were calculated, and strains with high degradation rates were screened. S3. Strain identification: Genomic DNA was extracted from the strain, and PCR amplification was performed using universal primers 27F and 1492R for the bacterial 16S rRNA gene. The product was sequenced, and the sequence was compared with the NCBI database using BLAST. Combined with physiological and biochemical characteristics, the target strain G-1 with 100% homology to Bacillus hygroscopicus was obtained.

[0086] This application also proposes a method for preparing a feed additive that degrades vomitoxin, such as... Figure 1 As shown, it includes the following steps: S1. Seed culture preparation: Bacillus hygroscopicus strain G-1 was inoculated into a shake flask containing NB medium and cultured with shaking at 37℃ and 180 rpm for 22 h to obtain seed culture; Understandably, the shake-flask seed culture in this embodiment aims to achieve the initial activation and proliferation of strain G-1 from its storage state to a highly active growth state. Strict control of the culture temperature and shaking rate provides optimal growth kinetics for the cells, ensuring a metabolically vigorous and highly pure seed culture. This step is fundamental to the successful large-scale fermentation subsequent; a high-quality seed culture significantly shortens the fermentation lag period, ensuring the fermentation broth ultimately reaches the preset high viable cell count, providing a source guarantee for the preparation of highly efficient detoxified products.

[0087] S2. Fermentation Culture: The seed culture was inoculated into a fermenter containing fermentation medium at a volume of 10%. Fermentation was carried out at 37℃, 180 rpm, and an aeration rate of 1.0 vvm for 48 h to obtain the fermentation broth. The effective viable cell count of the fermentation broth was not less than 5.0 × 10⁻⁶. 9 CFU / mL; Understandably, this application embodiment uses a fermenter for large-scale cultivation. By precisely controlling the inoculum size, temperature, stirring rate, and aeration rate, an optimized environment for the large-scale, high-density growth of *Bacillus hygroscopicus* G-1 is created. An appropriate inoculum size facilitates the rapid establishment of growth dominance, while adequate aeration and stirring ensure a uniform supply of dissolved oxygen and nutrients, thereby maximizing the accumulation of bacterial biomass and metabolites. This step is crucial for obtaining a high-concentration, high-activity fermentation broth, directly determining the effective dosage and degradation potential of the functional strains in the final additive.

[0088] S3. Carrier pretreatment: The solid carrier is crushed by a pulverizer and passed through an 80-mesh sieve. The material on the sieve is returned to be crushed again. The moisture content is adjusted to 15%, and the carrier is sterilized by moist heat at 121℃ for 30 min. It is then cooled to room temperature for use to obtain the pretreated solid carrier. It is understood that the pretreatment of the solid carrier in this embodiment, including pulverization, sieving, water adjustment, and sterilization, aims to create a suitable microenvironment for bacterial adsorption and survival. Controlling the carrier particle size and water content optimizes its physical structure, improving its compatibility and carrying capacity with the liquid fermentation broth. Strict moist heat sterilization completely eliminates any contaminating bacteria carried by the carrier itself, preventing them from contaminating the product and competing with functional strains. This step ensures the purity and applicability of the carrier as a bacterial "warehouse," and is a crucial pretreatment for preparing stable and homogeneous solid additives.

[0089] S4. Mixing and post-processing: The fermentation broth and the pretreated solid carrier are mixed at a weight ratio of 30%:70%, and stirred at 50 rpm for 40 min using a twin-screw mixer until the coefficient of variation of the mixing uniformity is ≤10%. The uniformly mixed material is then spray-dried at 50℃ until the moisture content is ≤12% to obtain a powdered feed additive.

[0090] It is understood that the embodiments of this application, by controlling the mixing ratio of bacterial solution and carrier, employing mechanical mixing at specific speeds and times, and finally performing gentle drying, aim to achieve efficient, uniform loading and stable solidification of functional strains on the carrier. Precise mixing ratios are fundamental to balancing product efficacy and cost; gentle yet thorough mixing ensures uniform bacterial dispersion; the subsequent low-temperature or rapid drying process aims to remove excess moisture for preservation while minimizing thermal damage to bacterial activity. This step ultimately determines the product's physical properties, viable cell survival rate, and storage stability, and is the final step in obtaining a high-quality, directly applicable powdered feed additive.

[0091] In step S1, the NB culture medium consists of: 10 g / L peptone, 3 g / L beef extract, 5 g / L sodium chloride, water as solvent, and a pH of 7.0.

[0092] In step S4, the drying process is spray drying.

[0093] This application also proposes the application of a feed additive that degrades vomitoxin, used to reduce the vomitoxin content in grains or feed and inhibit mold growth.

[0094] Comparative Example 1 This comparative example provides a feed additive for degrading vomitoxin and its application. The only difference between this example and Example 1 is that it does not contain Bacillus hygroscopicus G-1 bacterial solution. Instead, the amount of Bacillus hygroscopicus G-1 bacterial solution is replaced with an equal weight of sterile water or inactivated fermentation broth. The remaining components, component contents, and application process are the same as in Example 1.

[0095] Comparative Example 2 This comparative example provides a feed additive for degrading vomitoxin and its application. The only difference between this example and Example 1 is that the Bacillus hygroscopicus G-1 bacterial solution is replaced with an equal volume of Bacillus subtilis ATCC6051 fermentation broth with the same number of viable bacteria. The other components, component contents, preparation methods and application processes are the same as in Example 1.

[0096] Comparative Example 3 This comparative example provides a feed additive for degrading vomitoxin and its application. The only difference between this example and Example 1 is that the solid carrier is just ordinary wheat bran without any pretreatment. The other components, component contents, preparation methods and application processes are the same as in Example 1.

[0097] Performance testing Strain identification, degradation efficiency and environmental adaptability testing To clarify the taxonomic position of the core strain of this application and to systematically evaluate its basic ability to degrade vomitoxin (DON) and its environmental tolerance, the following tests were conducted. Figure 2 The image shows the colony and cell morphology of strain G-1, which exhibits typical Bacillus characteristics. Figure 3 A phylogenetic tree constructed based on the 16S rRNA gene sequence confirmed that strain G-1 is related to Bacillus hygroscopicus (Bacillus hygroscopicus). Bacillus altitudinis The homology is 100%.

[0098] The degradation kinetics of DON by strain G-1 under pure culture conditions were determined, showing a degradation rate of 85.3% within 48 hours. Simultaneously, the growth response of the strain was quantified by measuring its growth curves under different temperature and pH conditions to assess its environmental adaptability. Related results are as follows: Figure 4 and Figure 5 As shown: Figure 4 The effect of temperature on the growth of strain G-1 Figure 5 The effect of pH on the growth of strain G-1 is shown. Test data for all examples and comparative examples are summarized in Table 1.

[0099] Table 1. Data on strain identification, degradation efficiency, and environmental adaptability.

[0100] As shown in Table 1, the *Bacillus hygroscopicus* G-1 strain obtained in this application (Examples 1-5) exhibits superior DON degradation efficiency and environmental adaptability. Regarding degradation efficiency, strain G-1 achieved a DON degradation rate of up to 85.3% within 48 hours, while Comparative Example 2, *Bacillus subtilis*, only achieved 32.5%. This significant difference stems from the fact that strain G-1 can secrete specific deepoxidases and oxidoreductases with high affinity for key toxic sites such as the C12, 13-epoxy rings and C3-hydroxyl groups of the DON molecule. These enzymes can precisely catalyze the conversion of DON into low-toxicity or non-toxic deepoxidized products such as DOM-1, or further hydroxylate them into stable derivatives. These products are then further assimilated through the bacterial secondary metabolic system, thus achieving a complete pathway from toxin decomposition to complete conversion, eliminating the accumulation of intermediate toxic substances and the risk of toxin regeneration. In terms of environmental adaptability, strain G-1 maintained a relative growth rate of 75.2% at 60℃ and 52.1% in an acidic environment of pH 5.0. This is attributed to the adjustable composition of its cell membrane lipids and the efficient expression of protective mechanisms such as heat shock proteins and acid-base tolerance enzymes. This wide temperature range (27-67℃) and wide pH tolerance (5.0-9.0) characteristics allow it to withstand the high-temperature shock (80-90℃) during feed pelleting for short periods and maintain metabolic activity in the acidic environment of the stomach to the alkaline environment of the intestines, fundamentally solving the technical problem of easy inactivation of most strains under extreme processing and application conditions. In summary, the core strain G-1 of this application, through its efficient and specific enzymatic degradation system and strong intrinsic stress resistance mechanism, achieves efficient, thorough, and stable biological detoxification, laying a solid microbiological foundation for subsequent product development.

[0101] Test on the combined effects of detoxification and mold prevention of feed additives in simulated moldy feed To evaluate the application effect of the feed additives of this application in actual moldy feed matrix, fermentation tests were conducted on high-moisture moldy bran (initial DON 1.2 mg / kg) with the additives prepared in Examples 1-5 and Comparative Examples 1-3 respectively. Figure 6 The curves showing the changes in the content of vomitoxin (DON) in the moldy bran treated in Example 3 and Comparative Example 1 during fermentation are shown. Figure 7 The table shows the dynamic changes in microbial community and pH during fermentation in Example 3 and Comparative Example 1. (A) represents the change in bran pH, (B) the change in total bacterial count, (C) the change in mold count, and (D) the change in lactic acid bacteria count. The experiment monitored DON residue and mold inhibition at the fermentation endpoint (7 days). Key data are summarized in Table 2. Table 2. Detoxification and Anti-mold Effects of Additives on Moldy Wheat Bran

[0102] Combined with Table 2 Figure 6 and Figure 7 Analysis shows that the additives prepared in Examples 1-5 of this application exhibit excellent dual effects of detoxification and mold prevention in complex feed systems. Regarding detoxification, all examples reduced DON content by approximately 90% (degradation rate 88.3%-91.7%), which is consistent with the high-efficiency degradation capability under pure culture conditions. Figure 6 As shown, the toxin content in Example 3 group decreased rapidly and continuously during the later stages of fermentation, while that in Comparative Example 1 group continuously increased, providing direct evidence of the necessity of the core strain of this invention. Regarding mold prevention, the additive inhibited the number of molds by more than 91% and rapidly lowered the system pH to below 4.7. This "acid-lowering and mold-inhibiting" effect stems from multiple mechanisms: firstly, strain G-1 rapidly colonizes due to its vigorous growth, competitively consuming resources; secondly, the large amount of organic acids produced by its metabolism not only directly lowers the environmental pH, but its derived antibacterial substances also destroy the mold cell membrane. (See Table 2 and...) Figure 7 As shown, Comparative Example 1 (without live bacteria) and Comparative Example 2 (alternative strain) showed no difference in effect from the blank control due to the lack of core functional strains. The effect of Comparative Example 3 (carrier without pretreatment) was significantly weakened, highlighting the importance of the carrier pretreatment process. In summary, the feed additive of this application achieves comprehensive control from degrading existing toxins to inhibiting mold regeneration through a dual synergistic mechanism of "efficient enzymatic detoxification" and "rapid acid production to inhibit mold growth."

[0103] Processing tolerance and storage stability tests of feed additive products To ensure that the products of this application can withstand feed industry processing and meet commercial storage requirements, high-temperature pelleting treatment was simulated and a long-term accelerated storage test was conducted. Each product was subjected to dry heat treatment at 85°C for 3 minutes to simulate pelleting conditions. Simultaneously, an accelerated storage test was conducted for 6 months under standard conditions of 25°C and 60% relative humidity. Viable cell count and DON degradation activity retention were periodically monitored, and the results are as follows: Table 3 Processing tolerance and storage stability of feed additives

[0104] As shown in Table 3, the products of Examples 1-5 of this application exhibit good processing tolerance and long-term storage stability, which directly reflects the comprehensive advantages of the complete technical solution. After a short-term heat treatment at 85℃ simulating feed pelleting, the viable cell survival rate and degradation activity retention rate of all examples were higher than 84%; after 6 months of accelerated storage, both key indicators remained above 79%. This excellent stability is primarily attributed to the strong stress resistance of strain G-1 itself, whose heat shock proteins and other protective mechanisms effectively alleviate high-temperature damage; secondly, the complete preparation method used in the examples, especially the pretreatment of the solid carrier through crushing, sieving, moisture content adjustment, and moist heat sterilization, and the final use of low-temperature drying or spray drying at 40-50℃, together construct a solid microenvironment conducive to the adsorption and colonization of bacteria (especially spores) and providing physical protection. The carrier not only serves as a physical support, but its porous structure can also adsorb metabolites and moisture, providing stable protection for the bacteria during storage, thereby achieving slow release and long-lasting effects. In contrast, Comparative Example 2 (alternative strain) showed a significant decrease in stability (only about 60% activity retention after 6 months) due to the poor heat resistance of the strain itself, and Comparative Example 3 (incomplete process) showed a significant decrease due to the failure of the carrier to provide effective protection. This fully demonstrates that using a specific Bacillus cereus strain G-1, coupled with its complete and specific preparation process, is indispensable for obtaining a commercial feed additive product that can withstand actual processing conditions and maintain high activity throughout its shelf life. In summary, the product of this application successfully combines the biological characteristics of a highly efficient strain with an optimized formulation process, ensuring functional reliability throughout the entire chain from factory production to end-use. In summary, the Bacillus hygroscopicus G-1 and its feed additives provided in Examples 1-5 of this application, through systematic performance testing, fully demonstrate their comprehensive advantages in efficiently degrading vomitoxin (DON), inhibiting mold growth, broad environmental adaptability, and excellent product stability. Test data shows that strain G-1 is the core and key to achieving efficient and thorough biological detoxification, and its performance significantly surpasses that of the conventional strains used in the comparative examples. Simultaneously, a complete and specific production process (including refined carrier pretreatment) is a necessary condition for ensuring the realization and maintenance of this excellent effect. The strong environmental tolerance and long-term storage stability exhibited by the final product ensure that it can fully meet the stringent requirements of industrial feed processing, storage, and use. The systematic and regular significant differences between the examples and the comparative examples in all key indicators strongly support the inventiveness, practicality, and significant technological advancement of this invention.

[0105] According to the embodiments of this application, a feed additive for degrading vomitoxin and its application are proposed. Through screening, the *Bacillus glomeratus* strain G-1, combined with an optimized solid carrier and preparation process, achieves efficient and complete biodegradation of vomitoxin, while also exhibiting significant anti-mold properties. The core advantages of this additive are: strain G-1 can secrete a specific degrading enzyme system, achieving a degradation rate of over 85% for DON within 48 hours, and can convert the toxin into non-toxic metabolites, eliminating secondary pollution; it possesses a wide temperature range of 27-67℃ and a pH tolerance range of 5.0-9.0, adapting to the high temperatures of feed processing and the animal digestive tract environment; during application, it not only rapidly reduces toxin content but also effectively inhibits mold growth through acid production and nutrient competition, blocking toxin regeneration at the source; furthermore, the product prepared by a specific process exhibits good processing tolerance and storage stability. This invention provides a safe, efficient, and stable biological detoxification solution, thereby solving the problems of limited efficiency, unstable activity in complex environments, and lack of ability to inhibit mold regeneration in existing degrading strains.

[0106] Although embodiments of this application have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and spirit of this application, the scope of which is defined by the appended claims and their equivalents.

[0107] The present application and its embodiments have been described above. This description is not restrictive, and the actual application is not limited thereto. In conclusion, if a person skilled in the art is inspired by this description and designs a similar structure and embodiment without departing from the spirit of this application, such design should fall within the protection scope of this application.

Claims

1. A feed additive for degrading vomitoxin, characterized in that, Its composition by weight percentage is as follows: Highland Bacillus G-1 bacterial suspension: 10-0%; Solid carrier: 70-90%.

2. The feed additive for degrading vomitoxin according to claim 1, characterized in that, The effective viable count of the *Bacillus hygroscopicus* strain G-1 in the bacterial suspension is not less than 5.0 × 10⁻⁶. 9 CFU / mL.

3. The feed additive for degrading vomitoxin according to claim 1, characterized in that, The Bacillus subtilis strain G-1 can grow and degrade vomitoxin under conditions of 27-67℃ and pH 5.0-9.

0.

4. The feed additive for degrading vomitoxin according to claim 1, characterized in that, The solid carrier is one or more of the following: wheat bran, corn flour, rice husk powder, zeolite powder, montmorillonite, or diatomaceous earth.

5. A method for screening and obtaining the Bacillus subtilis strain G-1 as described in claim 1, characterized in that, Includes the following steps: S1. Separation and purification: Take 10 g of the sample containing microorganisms and place it in 100 mL of sterile water. Shake at 37℃ and 180 r / min for 1 h. Take the supernatant and perform a 10-fold serial dilution. Select 10 g of the supernatant for further dilution. -4 Up to 10 -7 Diluted bacterial suspensions were spread on NA plates and incubated at 37°C for 24 h. Single colonies were picked and subjected to at least three streak purification cultures to obtain pure cultures. S2. Initial screening of degradation performance: The pure culture was inoculated into NB medium and cultured at 37℃ and 180 r / min for 24 h. 950 μL of fermentation supernatant was mixed with 50 μL of vomitoxin standard solution with a concentration of 100 μg / mL and co-cultured at 60℃ for 72 h. The absorbance was measured at wavelengths of 450 / 630 nm using an enzyme-linked immunosorbent assay kit. The residual amount of vomitoxin and the degradation rate were calculated, and strains with high degradation rates were screened. S3. Strain identification: Genomic DNA was extracted from the strain, and PCR amplification was performed using the universal primers 27F and 1492R for the bacterial 16S rRNA gene. The product was sequenced and the sequence was compared with the NCBI database using BLAST. Combined with physiological and biochemical characteristics, the target strain G-1 with 100% homology to Bacillus hygroscopicus was obtained. A method for preparing a feed additive that degrades vomitoxin as described in any one of claims 1-4, characterized by comprising the following steps: S1. Seed culture preparation: Bacillus hygroscopicus strain G-1 was inoculated into a shake flask containing NB medium and cultured with shaking at 37℃ and 180 rpm for 14-24 h to obtain seed culture; S2. Fermentation Culture: The seed culture is inoculated into a fermenter containing fermentation medium at a volume inoculation rate of 5-10%, and fermented at 37℃, 180 rpm, and an aeration rate of 0.5-1.0 vvm for 24-48 h to obtain the fermentation broth. The effective viable cell count of the fermentation broth is not less than 5.0 × 10⁻⁶. 9 CFU / mL. 6.S3. Carrier pretreatment: The solid carrier is crushed by a pulverizer and then passed through an 80-mesh sieve. The material on the sieve is returned to be crushed again and then sterilized by moist heat at 105-121℃ for 15-30 min. After cooling to room temperature, the pretreated solid carrier is obtained. S4. Mixing and post-processing: The fermentation broth and the pretreated solid carrier are mixed at a weight ratio of 10-30%:70-90%, and stirred at 30-50 rpm for 20-40 min using a twin-screw mixer until the coefficient of variation of the mixing uniformity is ≤10%. The uniformly mixed material is then dried at 40-50℃ until the moisture content is ≤12% to obtain a powdered feed additive.

7. The method for preparing a feed additive that degrades vomitoxin according to claim 6, characterized in that, In step S1, the NB culture medium has the following composition: 10 g / L peptone, 3 g / L beef extract, 5 g / L sodium chloride, water as solvent, and pH value of 7.0-7.

2.

8. The method for preparing a feed additive that degrades vomitoxin according to claim 6, characterized in that, In step S3, the solid carrier is wheat bran, and its pretreatment includes crushing to a particle size ≤0.5 mm and adjusting the moisture content to 10-15%.

9. In the method for preparing a feed additive that degrades vomitoxin according to claim 6, in step S4, the drying is performed at a low temperature of 40-50°C or by spray drying.

10. The application of a feed additive for degrading vomitoxin as described in any one of claims 1-4, used to reduce the vomitoxin content in grains or feed and inhibit mold growth.

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