Bacillus coagulans and applications thereof

Abstract

The application belongs to the technical field of functional microorganism screening and application, and particularly relates to a Bacillus coagulans and application thereof. The Bacillus coagulans is isolated from a fecal sample of a healthy pig, and has been preserved in the China Center Type Culture Collection Center of Wuhan University in Wuhan, China on December 26, 2024, with a preservation number of CCTCC NO: M20242912. The strain does not produce hemolysin, cannot dissolve blood cells, is sensitive to various antibiotics, has good safety, has excellent heat resistance, gastric acid resistance and bile salt resistance characteristics, has high organic acid yield, can efficiently remove zearalenone toxin, can be widely applied in livestock and poultry breeding as a feed additive or mycotoxin detoxification agent, can significantly promote the growth of breeding animals, improve the immune performance and antioxidant performance of the animals, improve the intestinal flora structure, maintain the intestinal health, and has a wide application prospect.

Description

Technical Field

[0001] This invention belongs to the field of functional microbial screening and application technology, specifically relating to a Bacillus coagulans and its application. Background Technology

[0002] In modern animal husbandry, the long-term and irrational use of antibiotics not only threatens animal health but also poses a potential risk to human public health and safety. Therefore, restrictions on the use of antibiotics in feed are becoming increasingly stringent globally, making the search for antibiotic alternatives urgent.

[0003] Furthermore, the problem of mycotoxin contamination in feed ingredients is becoming increasingly serious in today's feed industry and livestock production. Feed is highly susceptible to mold contamination during planting, storage, and processing, leading to the production of various mycotoxins. Zearalenone is a common and highly hazardous mycotoxin found in feed. In livestock production, when animals ingest feed contaminated with zearalenone, it not only severely damages the quality and nutritional value of the feed but also poses potential threats to animal growth performance, reproductive function, and human health. Traditional methods for removing zearalenone from feed mainly include physical adsorption and chemical treatment. However, physical adsorption has limited effectiveness, and chemical treatment may introduce new chemical residues, posing a potential threat to animal health and food safety. Moreover, these methods often require complex equipment and processes, resulting in high costs and making large-scale application in actual production difficult.

[0004] Currently, biological detoxification is the most common method for removing zearalenone toxins from zearalenone. This method primarily utilizes microbial enzymes to break down mycotoxins or adsorbs the toxins onto the cell walls of microorganisms, thus achieving effective detoxification. This approach offers advantages such as high efficiency, environmental friendliness, and low cost, and can be widely applied in the food, feed, and agricultural sectors, significantly reducing the harm of mycotoxins to humans and animals. Therefore, screening for highly efficient mycotoxin-degrading microbial strains has become a research hotspot in recent years. Summary of the Invention

[0005] The purpose of this invention is to provide a Bacillus coagulans strain and its applications. This Bacillus coagulans strain exhibits excellent heat resistance, gastric acid resistance, and bile salt resistance; it also has a high organic acid yield and can efficiently degrade zearalenone toxin, showing broad application prospects.

[0006] In one aspect, this invention provides a Bacillus coagulans strain, named WL-64, which was deposited on December 26, 2024, at the China Center for Type Culture Collection, Wuhan University, Wuhan, China, with accession number CCTCC NO: M20242912.

[0007] The 16S rDNA sequence of the Bacillus coagulans strain WL-64 is SEQ ID NO: 1.

[0008] The present invention also provides the application of the aforementioned Bacillus coagulans WL-64 in feed or food production.

[0009] The present invention also provides the application of the aforementioned Bacillus coagulans WL-64 in the production of mycotoxin detoxifiers.

[0010] The mycotoxin is any one or more of zearalenone, aflatoxin, fumonisin, ochratoxin, T-2 toxin, and vomitoxin.

[0011] The present invention also provides a microbial preparation comprising the Bacillus coagulans WL-64.

[0012] The present invention also provides the application of the aforementioned microbial preparation in feed or food production.

[0013] The Bacillus coagulans WL-64 provided by this invention is non-hemolytic, sensitive to kanamycin, gentamicin, ampicillin, erythromycin, azithromycin, and tetracycline, and has high safety; it also has extremely strong heat resistance, gastric acid resistance, and bile salt resistance.

[0014] Bacillus coagulans WL-64 can metabolize high levels of organic acids. After 24 hours of fermentation, the yields of lactic acid, formic acid, acetic acid, and propionic acid in its supernatant reached 15.04 g / L, 0.9 g / L, 9.2 g / L, and 10.3 g / L, respectively.

[0015] Bacillus coagulans WL-64 exhibited significant inhibitory effects against Escherichia coli K88, Staphylococcus aureus, Salmonella pullorum (chicken), Salmonella suis, Clostridium perfringens (chicken), Clostridium perfringens (suis), Haemophilus parasuis, and Streptococcus suis SS-2. Among these, Bacillus coagulans WL-64 showed the best inhibitory effect against Salmonella pullorum (chicken), with an inhibition zone diameter reaching 23.5 mm.

[0016] Bacillus coagulans WL-64 has a strong antioxidant capacity. Its fermentation broth scavenged 85.3% and 58.3% of DPPH free radicals and hydroxyl free radicals, respectively, while its bacterial suspension scavenged 62.9% and 50.7% of DPPH free radicals and hydroxyl free radicals, respectively, achieving unexpected technical results.

[0017] Bacillus coagulans WL-64 can efficiently remove zearalenone, achieving a removal rate of 89.7% for zearalenone in liquid culture medium. This strain can be directly applied to detoxify feed ingredients. After 24 hours of treatment, the concentrations of zearalenone in corn flour, soybean meal, wheat bran, and cottonseed meal decreased by 90.0%, 73.2%, 68%, and 86.3%, respectively, demonstrating significant detoxification effects.

[0018] Bacillus coagulans WL-64 can be widely used as an additive or mycotoxin detoxifier in food or feed production. It can significantly improve the body's immune and antioxidant properties, improve the intestinal flora structure, maintain intestinal health, and promote growth, showing broad application prospects. Attached Figure Description

[0019] Figure 1 This is a colony morphology diagram of Bacillus coagulans WL-64;

[0020] Figure 2 Microscopic morphological image of Bacillus coagulans WL-64;

[0021] Figure 3 The image shows the inhibitory effect of Bacillus coagulans WL-64 on Salmonella pullorum. Detailed Implementation

[0022] The following are specific embodiments of the present invention, which describe the technical solution of the present invention in detail, but are not intended to limit the scope of protection of the present invention.

[0023] Example 1: Screening of Bacillus coagulans WL-64

[0024] A 10g fecal sample from a healthy pig at a pig farm in Qingdao was added to a 250ml Erlenmeyer flask containing glass beads. 90mL of physiological saline was added and thoroughly mixed. The sample was then incubated at 80℃ for 10min to kill non-spore-forming bacteria. The sample was serially diluted, and 100μL of each appropriate serial dilution was spread onto MRS solid plates and incubated upside down at 45℃ for 48h. Single colonies isolated from multiple streaks were inoculated into MRS liquid medium and incubated at 45℃ for 24h. Microscopic examination of bacterial cell and spore morphology was performed. Five strains—WL-3, WL-8, WL-12, WL-38, and WL-64—were Gram-positive, and their spores were terminal. Further pH measurements of the fermentation broth for each of the five strains were conducted to screen for strains with high acid production.

[0025] The results showed that strain WL-64 had the strongest acid production capacity, and the pH dropped to 4.0 after 24 hours of culture at 45℃.

[0026] Example 2: Identification of Bacillus coagulans WL-64

[0027] 1. Morphological identification:

[0028] The WL-64 strain grew well on MRS solid medium, producing moist, white colonies with regular edges and smooth surfaces. Figure 1 Microscopic observation revealed that the bacteria were rod-shaped, occurring singly or in pairs, with terminal, oval-shaped spores. Figure 2 ).

[0029] 2. Physiological and biochemical identification:

[0030] Physiological and biochemical identification of the strain was performed, and the results are shown in Table 1. The strain met the physiological and biochemical characteristics of Bacillus coagulans and was preliminarily identified as Bacillus coagulans.

[0031] Table 1. Physiological and biochemical identification of strain WL-64

[0032]

[0033] Molecular biological identification: The genome of strain WL-64 was extracted. Using the genome as a template, PCR amplification was performed using universal primers 27F (5′-AGAGTTTGATCCTGGCTCAG-3′) and 1492R (5′-GGTTACCTTGTTACGACTT-3′) for bacterial 16S rDNA. The amplification conditions were: 98℃ for 3 min; 98℃ for 30 s, 55℃ for 30 s, 72℃ for 90 s, for 30 cycles; 72℃ for 10 min. The amplified positive fragments were purified and sent to Shanghai Bioengineering Co., Ltd. for sequencing.

[0034] The 16S rDNA sequence of strain WL-64 is shown in SEQ ID NO:1. Details are as follows:

[0035]

[0036] Homology comparison analysis of SEQ ID NO:1 with related sequences in NCBI showed that strain WL-64 is Bacillus coagulans, named Bacillus coagulans WL-64, and deposited on December 26, 2024 at the China Center for Type Culture Collection of Wuhan University, Wuhan, China, with accession number CCTCC NO: M20242912.

[0037] Example 3: Characteristic analysis of Bacillus coagulans WL-64

[0038] Bacillus coagulans strain WL-64, stored at -80℃, was streaked onto MRS solid medium and incubated at 45℃ for 48 h. Single colonies were picked and inoculated into MRS liquid medium and incubated at 45℃ for 17 h. Bacillus coagulans strain WL-64 was then transferred to fresh MRS liquid medium at a 10% inoculum size and incubated at 45℃ with shaking for 24 h. The bacterial cells were obtained by centrifugation at 4000 rpm for 5 min. The bacterial cells were washed twice with physiological saline and then resuspended in physiological saline to obtain a bacterial suspension.

[0039] 3.1 Hemolytic test:

[0040] Prepare TBS basal medium, autoclave at 121℃ for 15 min, and after the medium cools to 50℃, add 5% sterile defibrinated sheep blood and mix well. Pour into plates; pick a single colony of Bacillus coagulans WL-64 and streak it onto the prepared blood cell plate. Incubate at 37℃ for 24 h and observe whether the test bacteria exhibit hemolysis.

[0041] The results showed that no lysate zones appeared, meaning that Bacillus coagulans WL-64 does not produce hemolysin and cannot lyse blood cells, indicating good safety.

[0042] 3.2 Antibiotic susceptibility testing:

[0043] The susceptibility of bacterial strains to antibiotics was determined using the agar diffusion disk method (susceptibility: inhibition zone greater than 20 mm). The susceptibility level of the strain to antibiotics was determined by the size of the inhibition zone.

[0044] Table 2 Antibiotic susceptibility test of Bacillus coagulans WL-64

[0045]

[0046] The results are shown in Table 2. Bacillus coagulans WL-64 was sensitive to kanamycin, gentamicin, ampicillin, erythromycin, azithromycin, and tetracycline, and had good biosafety.

[0047] 3.3 Tolerance to artificial gastric and intestinal fluids:

[0048] The bacterial suspension was added to simulated gastric fluid (pH=1.5, 2.0) and simulated intestinal fluid (0.3% bile salt, 0.5% bile salt) for 2 hours, respectively. Samples were taken for plate colony counting. The number of viable bacteria without gastric fluid or bile salt was used as a control, and the survival rate of the strain was calculated.

[0049] The results showed that Bacillus coagulans WL-64 had a survival rate of over 90% after treatment with artificial gastric and intestinal fluids, demonstrating extremely strong tolerance.

[0050] 3.4 Heat resistance:

[0051] When the Bacillus coagulans WL-64 bacterial suspension was treated at different temperatures of 70℃, 80℃, 85℃ and 90℃ for 10 min, the survival rate was above 90%, indicating that the strain has strong heat resistance.

[0052] 3.5 Determination of Organic Acid Yield:

[0053] After Bacillus coagulans WL-64 was cultured in MRS liquid medium at 45°C with shaking for 24 h, the supernatant was taken for determination of organic acid content.

[0054] The results showed that Bacillus coagulans WL-64 produced high levels of organic acids, with lactic acid, formic acid, acetic acid, and propionic acid yields reaching 15.04 g / L, 0.9 g / L, 9.2 g / L, and 10.3 g / L, respectively.

[0055] Example 4: Analysis of the antibacterial activity of Bacillus coagulans WL-64

[0056] The inhibitory effect of Bacillus coagulans WL-64 on common animal pathogens was detected using the Oxford cup method.

[0057] First, based on the type and requirements of the pathogen, select a suitable culture medium to prepare 10 6 Add 0.1 mL of a CFU / mL pathogen suspension to a suitable solid agar plate for pathogen growth. 6 CFU / mL of pathogenic bacterial suspension. Then place a 6mm Oxford cup in the center of the plate and add 50μL of a 10 CFU / mL solution to the Oxford cup. 6 A CFU / mL suspension of Bacillus coagulans WL-64 was incubated at 37°C for 24 h, and the diameter of the inhibition zone was measured. A control was prepared by inoculating 50 μL of MRS liquid medium. Each experiment was repeated three times.

[0058] Table 3. In vitro antibacterial test of Bacillus coagulans WL-64

[0059]

[0060] Table 3 shows that *Bacillus coagulans* WL-64 significantly inhibited the growth of *Escherichia coli* K88, *Staphylococcus aureus*, *Salmonella pullorum*, *Salmonella suis*, *Clostridium perfringens* (chicken-derived), *Clostridium perfringens* (swine-derived), *Haemophilus parasuis*, and *Streptococcus suis* SS-2. Among these, *Bacillus coagulans* WL-64 showed the best inhibitory effect against *Salmonella pullorum*, with an inhibition zone diameter of 23.5 mm. Figure 3 ).

[0061] Example 5: Determination of the in vitro antioxidant capacity of Bacillus coagulans WL-64

[0062] (1) DPPH free radical scavenging ability

[0063] Take 1 mL of Bacillus coagulans fermentation broth and bacterial suspension, respectively, and mix thoroughly with 1 mL of DPPH solution (0.4 mmol / L). Incubate at room temperature in the dark for 30 min. After the reaction, measure the absorbance (A) of the sample at 517 nm. 样品 An equal volume of physiological saline was used as a control to replace the sample; the absorbance of the control group was denoted as A. 对照 .

[0064] The DPPH free radical scavenging rate of the strain was calculated as follows:

[0065] Clearance rate (%) = (1 – A) 样品 / A 对照 )×100%.

[0066] (2) Hydroxyl radical scavenging ability

[0067] 200 μL of bacterial suspension and fermentation broth were thoroughly mixed with 200 μL of 2.5 mmol / L 1,10-phenanthroline, 200 μL of 2.5 mmol / L FeSO4 solution, 200 μL of 0.02 mmol / L PBS, and 200 μL of 20 mmol / L H2O2. The mixtures were then incubated in a water bath at 45 °C for 1 h. The absorbance (A) of the samples was measured at 536 nm. a .

[0068] The formula for calculating hydroxyl radical scavenging rate is as follows:

[0069] Hydroxyl radical scavenging rate = (A a -A b ) / (A c -A b )×100%.

[0070] Among them, A b To replace the test sample with an equal volume of deionized water, A c The sample and H2O2 were replaced with an equal volume of deionized water.

[0071] The results showed that Bacillus coagulans WL-64 has a strong antioxidant capacity. Its fermentation broth scavenged 85.3% and 58.3% of DPPH free radicals and hydroxyl free radicals, respectively, while the bacterial suspension scavenged 62.9% and 50.7% of DPPH free radicals and hydroxyl free radicals, respectively, achieving unexpected technical results.

[0072] Example 6: Preparation of Bacillus coagulans WL-64 bacterial powder

[0073] 1. Preparation of Bacillus coagulans WL-64 fermentation broth

[0074] Single colonies of Bacillus coagulans WL-64 were picked and inoculated into MRS liquid medium and cultured at 45°C for 17 h. Bacillus coagulans WL-64 was then transferred to fresh MRS liquid medium at a 10% inoculum and cultured at 45°C with shaking for 24 h as seed culture.

[0075] The Bacillus coagulans seed culture of WL-64 was transferred to the fermentation medium at a volume ratio of 5%. The fermentation formula was as follows: soybean meal 10.0 g / L, corn steep liquor 10.0 g / L, corn starch 10.0 g / L, dipotassium hydrogen phosphate 2.0 g / L, magnesium sulfate 0.58 g / L, and manganese sulfate 0.25 g / L. The culture was carried out at 45℃ and 200 rpm for 20-24 h. Microscopic examination showed that the Bacillus coagulans WL-64 morphology was normal, with no contaminating bacteria, and the spore count was 90% or higher, at which point fermentation was terminated. After fermentation, the temperature was raised to 60℃ and maintained for 10 min to obtain the Bacillus coagulans WL-64 fermentation broth. The spore count of Bacillus coagulans WL-64 in the fermentation broth was 9.0 × 10⁶. 9 CFU / mL, with a spore rate of 90% or higher.

[0076] 2. Preparation of Bacillus coagulans WL-64 bacterial powder

[0077] The fermentation broth of Bacillus coagulans WL-64 was thoroughly mixed with a protectant (10% corn starch, 5% sucrose, 2% gum arabic, 1% vitamin C, and 0.5% L-monosodium glutamate) to obtain a spray mixture. A centrifugal spray dryer was used for spray drying, controlling the inlet air temperature at 150℃ and the outlet air temperature at 70℃. The bacterial powder yield reached 85% or higher, and the number of Bacillus coagulans spores in the bacterial powder was 10-1. 11 CFU / g.

[0078] Example 7: Degradation of zearalenone toxin by Bacillus coagulans WL-64

[0079] 1. Detoxification effect of Bacillus coagulans WL-64 in liquid culture medium

[0080] Take 1g of bacteria as 10 11CFU / g of Bacillus coagulans WL-64 inoculum powder was inoculated into 100 mL of MRS liquid medium, and zearalenone was added to a final concentration of 5 μg / mL. The medium was incubated at 45℃ and 200 rpm for 6 h, 12 h, 18 h, 24 h, and 32 h, respectively. The concentration of residual zearalenone in the medium was determined using a zearalenone ELISA kit. MRS liquid medium containing only zearalenone and no inoculation powder served as a control.

[0081] Zearalenone removal rate (%) = (zearalenone content in control group - zearalenone content in experimental group) / zearalenone content in control group × 100%.

[0082] The results showed that within 6-24 hours of cultivation, the removal rate of zearalenone by Bacillus coagulans WL-64 gradually increased with the extension of cultivation time. The removal effect of zearalenone was strongest at 24 hours, with a removal rate of 89.7%, achieving unexpected technical results.

[0083] 2. Detoxification effect of Bacillus coagulans WL-64 on feed ingredients corn flour, soybean meal, wheat bran, and cottonseed meal.

[0084] Take 1g of bacteria as 10 11 CFU / g of Bacillus coagulans WL-64 powder was inoculated into 100g of feed ingredients contaminated with zearalenone, including corn flour, soybean meal, wheat bran, and cottonseed meal. The mixture was thoroughly mixed and incubated at 45℃ for 24h. Feed ingredients not inoculated with Bacillus coagulans WL-64 served as a control.

[0085] 1g of feed ingredients from both the experimental and control groups were pulverized and passed through a 20-mesh sieve. The pulverized feed ingredients were placed in Erlenmeyer flasks, and 25mL of a methanol-water (6:4) mixture was added. The mixture was shaken for 15 minutes, filtered, and the filtrate was collected and appropriately diluted as the test solution. The zearalenone toxin content in the test solution was determined using a zearalenone enzyme-linked immunosorbent assay (ELISA) kit.

[0086] The results showed that after 24 hours of fermentation in corn flour, soybean meal, wheat bran, and cottonseed meal contaminated with zearalenone by Bacillus coagulans WL-64, the concentration of zearalenone decreased by 90.0%, 73.2%, 68%, and 86.3%, respectively, indicating that Bacillus coagulans WL-64 has a significant removal effect on zearalenone.

[0087] Example 8: Application of Bacillus coagulans WL-64 in broiler farming

[0088] The feeding trial was conducted at a chicken farm in Qingdao, Shandong Province. Three hundred 7-day-old yellow-feathered broilers were randomly divided into two groups, with five replicates per group and 30 birds per replicate. The experimental group received a basal diet containing 10%... 8 The study added CFU / kg of Bacillus coagulans WL-64 bacterial powder, while the control group was fed a basal diet. The experiment lasted for 45 days, with free access to food and water, and immunization was carried out according to the standard immunization schedule.

[0089] 1. Growth performance testing

[0090] Daily feed intake was recorded for each replicate during the experiment. Feed was taken after a 12-hour fast before the start and end of the experiment, and average daily feed intake, average daily weight gain, and feed conversion ratio were calculated.

[0091] Average daily feed intake = Total feed consumption / Number of days in the experiment.

[0092] Average daily weight gain = (final weight - initial weight) / trial days.

[0093] Feed conversion ratio = average daily feed intake / average daily weight gain.

[0094] The results showed that, compared with the control group, the experimental group of broilers had an average daily weight gain and average daily feed intake that increased by 35.5% and 17.7%, respectively, while the feed conversion ratio decreased by 13%.

[0095] 2. Immune organ index

[0096] At 52 days of age, after fasting for 12 hours, one chicken close to the average weight was selected from each replicate for slaughter. The spleen, bursa of Fabricius, and thymus were removed, weighed, and the indices of each immune organ were calculated.

[0097] Immune organ index (%) = (weight of immune organs / body weight) × 100%.

[0098] The results showed that, compared with the control group, the spleen index, bursa of Fabricius index, and thymus index of the experimental group broilers increased by 50.2%, 35.9%, and 64.2%, respectively. This demonstrates that the Bacillus coagulans WL-64 provided by this invention can significantly improve the immune function of broilers.

[0099] 3. Immune function

[0100] After the experiment, blood was collected from the subwing vein of 52-day-old broilers and centrifuged at 4000 r / min for 10 min to obtain serum. The levels of immunoglobulin A (IgA), immunoglobulin G (IgG), and immunoglobulin M (IgM) in the serum were determined using an ELISA kit from Nanjing Jiancheng Bioengineering Institute.

[0101] The results showed that, compared with the control group, the levels of IgA and IgM in the serum of broilers increased by 32.5% and 29.7%, respectively, while the IgG level did not change significantly. This indicates that Bacillus coagulans WL-64 can improve the immunity of yellow-feathered broilers to a certain extent and promote their growth and development.

[0102] 4. Antioxidant function

[0103] The contents of glutathione peroxidase (GSH-PX), total superoxide dismutase (T-SOD), total antioxidant capacity (T-AOC), and malondialdehyde (MDA) in broiler serum were determined using a kit from Nanjing Jiancheng Bioengineering Institute.

[0104] The results showed that, compared with the control group, the activities of T-SOD, GSH-Px, and T-AOC in the serum of broilers fed with Bacillus coagulans WL-64 increased by 32.0%, 25.7%, and 28.9%, respectively, while the serum MDA content decreased by 23.8%. This indicates that Bacillus coagulans WL-64 can significantly improve the antioxidant capacity of broilers.

[0105] 5. Effects of Bacillus coagulans WL-64 inoculum on cecal flora

[0106] Cecum were collected from broiler chickens in the experimental and control groups respectively. 1g of cecum was added to 9ml of sterile physiological saline and shaken well. The mixture was then diluted 10 times stepwise. The appropriate dilutions were inoculated into MacConkey medium and MRS solid medium respectively. After incubation at 37℃ for 24h and 48h respectively, Escherichia coli and lactic acid bacteria were counted.

[0107] The results showed that, compared with the control group, the number of Escherichia coli in the cecum of broilers fed with Bacillus coagulans WL-64 decreased by 32.1%, while the number of lactic acid bacteria increased by 25.7%. This indicates that Bacillus coagulans WL-64 can significantly improve the microbial community structure in the cecum of yellow-feathered broilers, which is beneficial to maintaining intestinal health.

[0108] In summary, the Bacillus coagulans WL-64 strain provided by this invention exhibits high biosafety and strong resistance to heat, gastric acid, and bile salts. This strain can metabolize high levels of organic acids and has significant inhibitory effects on various pathogens such as Escherichia coli K88 and Staphylococcus aureus. It can efficiently remove zearalenone, significantly improve the body's immune and antioxidant properties, improve intestinal flora structure, maintain intestinal health, and promote growth. It can be widely used as an additive or mycotoxin detoxifier in food or feed production, showing broad application prospects.

Claims

1. A Bacillus coagulans species, characterized in that, The preservation number of the Bacillus coagulans is CCTCC NO: M20242912.

2. The Bacillus coagulans as described in claim 1, characterized in that, The 16S rDNA sequence of the Bacillus coagulans is SEQ ID NO:

1.

3. The application of Bacillus coagulans as described in claim 1 in the preparation of food.

4. The application of Bacillus coagulans as described in claim 1 in the preparation of feed.

5. The application of Bacillus coagulans according to claim 1 in the preparation of mycotoxin detoxifiers, characterized in that, The mycotoxin in question is zearalenone.

6. A microbial preparation, characterized in that, The microbial preparation comprises Bacillus coagulans as described in claim 1.

7. The microbial preparation according to claim 6, characterized in that, The viable count of Bacillus coagulans in the microbial preparation is not less than 10. 9 CFU / g.

8. The use of the microbial preparation according to claim 6 or 7 in the preparation of feed.

9. The use of the microbial preparation according to claim 6 or 7 in the preparation of mycotoxin detoxifiers, characterized in that, The mycotoxin in question is zearalenone.

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