Bacillus and application of composition thereof

Abstract

The invention relates to application of a bacillus and a composition thereof. The invention discloses a bacillus strain and application thereof, and relates to the technical field of biological medicines and microbial agents. The invention relates to bacillus (Bacillus sp.), which is used for relieving colitis, is named as bacillus SLMG-002 strain, is preserved in China Center for Type Culture Collection, Wuhan University, on September 16, 2025, and has a preservation number of CCTCC M 20252231. The bacillus SLMG-002 strain can relieve colitis, the bacillus SLMG-002 strain and the bacillus SLMG-003 are combined for use to achieve a synergistic effect in the aspect of relieving intestinal inflammation, the effect of the bacillus SLMG-002 strain is better than that of single use of the bacillus SLMG-002, the bacillus SLMG-002 strain is used for reducing harm of colitis to humans or animals, and compared with other detoxification methods, the method is easy and convenient to operate, low in cost and high in safety. The invention provides a novel efficient solution for treating the inflammatory bowel disease.

Description

Technical Field

[0001] This invention belongs to the fields of biomedicine and microbial agents, specifically relating to the application of a strain of Bacillus and its composition. Background Technology

[0002] Gut homeostasis is a dynamic equilibrium formed by the interaction of the intestinal mucosa, immune barrier, and gut environment (including gut microbiota, nutrients, and metabolites). It ensures the effective absorption of nutrients and resists the invasion of harmful pathogens. However, when environmental changes, genetic factors, or unhealthy lifestyle habits disrupt this balance, gut microbiota imbalance occurs. The number of beneficial bacteria that originally coexist decreases, while harmful bacteria take the opportunity to proliferate, leading to impaired intestinal mucosal barrier function and overactivation of the immune system, ultimately resulting in inflammatory bowel disease (IBD).

[0003] Inflammatory bowel disease (IBD) is a chronic, relapsing gastrointestinal infection characterized by its high relapse rate. It primarily includes ulcerative colitis (UC) and Crohn's disease (CD). These diseases not only cause persistent symptoms such as abdominal pain, diarrhea, and rectal bleeding, severely impacting patients' quality of life, but also significantly increase the risk of developing intestinal tumors, posing a long-term threat to patients' health. Summary of the Invention

[0004] This invention provides the application of a Bacillus SLMG-002 strain and its composition, aiming to provide a method for rapidly relieving colitis, effectively reducing intestinal inflammation, repairing damaged intestinal mucosal tissue, and reducing the health hazards of inflammatory bowel disease to humans or animals. It was also found that the combined use of Bacillus SLMG-002 and Bacillus SLMG-003 strains has a synergistic effect in relieving intestinal inflammation, and its effect is superior to the use of Bacillus SLMG-002 alone.

[0005] The objective of this invention is achieved through the following technical solutions: A strain of Bacillus ( Bacillus sp. The strain, named Bacillus SLMG-002, was used to quickly relieve colitis and was deposited on September 16, 2025, at the China Center for Type Culture Collection (CCTCC) Wuhan University Collection Center, with accession number CCTCC M 20252231.

[0006] This invention also provides the application of a Bacillus SLMG-002 strain, which works synergistically through multiple pathways by regulating the balance of intestinal flora, repairing the intestinal mucosal barrier, and inhibiting intestinal inflammatory response, and can be used to prepare a bacterial agent that rapidly relieves colitis.

[0007] The present invention also provides a Bacillus composition comprising the above-mentioned Bacillus SLMG-002 strain and Bacillus SLMG-003 strain in a ratio of 3:7 to 7:3.

[0008] This invention also provides an application of a Bacillus composition, which works synergistically through multiple pathways by regulating the balance of intestinal flora, repairing the intestinal mucosal barrier, and inhibiting intestinal inflammatory response, and is used to prepare a bacterial agent that rapidly relieves colitis.

[0009] Preferably, the above-mentioned microbial agents include: liquid microbial agents and freeze-dried microbial agents.

[0010] Preferably, the above-mentioned liquid bacterial agent is prepared by inoculating Bacillus SLMG-002 strain into LB liquid medium and culturing it in a shaker at 37°C until the concentration reaches 4×10⁻⁶. 8 CFU / 50μL, packaged and refrigerated.

[0011] Preferably, the above-mentioned freeze-dried bacterial agent is prepared by inoculating Bacillus SLMG-002 strain into LB liquid medium and culturing it at 37°C in a shaker until the concentration reaches 4 × 10⁻⁶. 8 CFU / 50μL, freeze-dried, packaged, and refrigerated.

[0012] This invention also provides a method for applying Bacillus inoculant, wherein the above-mentioned Bacillus SLMG-002 inoculant is added to feed or drinking water at a dosage of 5 × 10⁻⁶. 8 CFU - 5 × 10⁹ CFU / kg of feed.

[0013] This invention also provides a method for applying the Bacillus composition, wherein the above-mentioned Bacillus composition is added to feed or drinking water at an amount of 5 × 10⁻⁶. 8 CFU-5×10 9 CFU / kg.

[0014] This invention utilizes traditional bacterial isolation methods to isolate a Bacillus strain SLMG-002 from the intestinal contents of animals subjected to L-fucose gavage. Morphological characteristics confirmed that this strain is Gram-positive; on LB agar, its colonies are white with a rough surface and serrated edges. The growth temperature range is 30-37℃, and the optimal pH for growth is 7.0. The 16S rRNA gene of this strain was amplified by PCR using primers 5'-AGAGTT TGA TCM TGG CTC AG-3' and 5'-CGG TTA CCT TGT TAC GAC TT-3'. The obtained sequence was submitted to the GeneBank public database with the sequence number SUB15727522. This sequence serves as the molecular characteristic for identifying the strain. Based on phylogenetic analysis of the 16S rRNA gene, the strain was identified as belonging to the genus Bacillus and named Bacillus SLMG-002. It is deposited at the China Center for Type Culture Collection (CCTCC) of Wuhan University with the accession number CCTCC M 20252231. This isolated Bacillus SLMG-002 strain demonstrated significant therapeutic effects with short-term use in a mouse model of DSS-induced colitis. It effectively alleviated the inflammatory response of DSS-induced colitis and salvaged intestinal damage caused by DSS, providing a new strategy for the treatment of inflammatory bowel disease and weaning diarrhea.

[0015] Bacillus strain SLMG-003, isolated from the intestinal contents of animals given L-fucose by gavage, was deposited on September 3, 2025, at the China Center for Type Culture Collection (CCCCC), Wuhan University, with accession number CCTCC M 20251956. This strain is protected by patent application number 2025113198771.

[0016] The advantages of this invention compared to existing technologies are: Bacillus SLMG-002 strain can alleviate colitis in a short period (within 10 days), reducing the harm caused by colitis to humans or animals. Compared to other detoxification methods, this method is simple to operate, low in cost, and highly safe. This Bacillus can be used in various forms such as oral preparations and feed additives, providing a new and highly effective solution for the treatment of inflammatory bowel disease.

[0017] Our isolated Bacillus strain SLMG-002 demonstrated significant therapeutic effects in DSS-induced colitis models in mice and weaned piglets. It effectively alleviated the inflammatory response of DSS-induced colitis and salvaged intestinal damage caused by DSS, providing a new strategy for the treatment of inflammatory bowel disease. The Bacillus SLMG-002 strain exerts its effects synergistically through multiple pathways, including regulating gut microbiota balance, repairing the intestinal mucosal barrier, inhibiting intestinal inflammatory responses, and producing beneficial metabolites such as short-chain fatty acids.

[0018] Compared to other treatments for inflammatory bowel disease, intervention using Bacillus SLMG-002 is simpler and easier to implement, has low short-term cost, and is highly safe. This Bacillus SLMG-002 can be applied through various routes, including oral formulations and feed additives, offering flexibility to adapt to different application scenarios and needs.

[0019] When Bacillus strain SLMG-002 is used in combination with the previously isolated Bacillus strain SLMG-003, it has a synergistic effect in relieving intestinal inflammation and is more effective than using either strain alone. Attached Figure Description

[0020] Figure 1 A statistical curve of mouse body weight during diarrhea modeling; Figure 2 Photograph of mouse intestines; Figure 3 This is a graph showing the statistical results of mouse intestinal length. Figure 4 Image showing the HE staining results of mouse ileum; Figure 5 Image showing the results of HE staining of mouse colon; Figure 6 A bar chart of white blood cell count in a complete blood count of piglets; Figure 7 A bar chart showing the concentration of immune factors in piglet serum; Figure 8 Histological images of the ileum and colon of piglets; Figure 9 Histological image of the colon of a piglet. Detailed Implementation

[0021] The method of the present invention will be further described below with reference to the accompanying drawings and specific embodiments.

[0022] Example 1 A strain of Bacillus ( Bacillus sp. This embodiment provides a Bacillus strain for rapid relief of colitis, named Bacillus SLMG-002, which was deposited on September 16, 2025 at the China Center for Type Culture Collection (CCTCC) Wuhan University Collection Center, with accession number CCTCC M 20252231.

[0023] Example 2 A method for preparing a liquid Bacillus SLMG-002 bacterial agent includes the following steps: Bacillus strain SLMG-002 was inoculated into LB liquid medium and cultured at 37°C in a shaker until the concentration reached 4 × 10⁻⁶. 8CFU / 50μL, packaged and refrigerated.

[0024] Example 3 A method for preparing a Bacillus SLMG-002 lyophilized bacterial agent includes the following steps: Bacillus strain SLMG-002 was inoculated into LB liquid medium and cultured at 37°C in a shaker until the concentration reached 4 × 10⁻⁶. 8 CFU / 50μL, freeze-dried, packaged, and refrigerated.

[0025] Example 4 A method for preparing a liquid Bacillus SLMG-003 bacterial agent includes the following steps: Bacillus strain SLMG-003 was inoculated into LB liquid medium and cultured at 37°C in a shaker until the concentration reached 4 × 10⁻⁶. 8 CFU / 50μL, packaged and refrigerated.

[0026] Example 5 A method for preparing a Bacillus SLMG-003 lyophilized bacterial agent includes the following steps: Bacillus strain SLMG-003 was inoculated into LB liquid medium and cultured at 37°C in a shaker until the concentration reached 4 × 10⁻⁶. 8 CFU / 50μL, freeze-dried, packaged, and refrigerated.

[0027] The DSS colitis model, due to its high similarity to human ulcerative colitis in pathological features, has become an important tool for studying the mechanisms and treatments of intestinal inflammation. Extensive experimental data show that DSS-induced mouse models exhibit significant weight loss, as intestinal inflammation affects nutrient absorption; simultaneously, intestinal tissue suffers severe damage, manifested as disruption of intestinal mucosal integrity and inflammatory cell infiltration.

[0028] Therefore, the embodiments of the present invention verify the application effect of Bacillus SLMG-002 strain through DSS modeling experiments. The embodiments of the present invention were only used for experiments on mice and pigs; in practical applications, other animals such as chickens, ducks, cattle, and sheep can also be selected.

[0029] Example 6 Mouse DSS modeling experiment and evaluation of diarrhea status and probiotic intervention in the DSS model. Fifteen 7-week-old male C57BL / 6 mice were selected and divided into a control group (n=5) and a DSS group (n=15). The control group mice had normal access to drinking water, while the DSS group mice freely ingested drinking water containing 2.5% DSS (MW: 36000~50000) to induce colitis. The drinking water was changed every two days for seven days. During this period, the mice were weighed daily, and their condition, anal sphincter function, and diarrhea were regularly observed. After the model was established, the DSS group mice were further divided into three groups: a self-healing group (n=5), a probiotic BS2 group (n=5), and a probiotic BS2 group (n=5) receiving 50 μL of approximately 4 × 10⁻⁶ DSS via gavage in addition to normal access to drinking water and feed. 8 CFU Bacillus sp. SLMG-002, once daily), 5 animals in the probiotic BS2+BS3 group (in addition to normal intake of drinking water and feed, the concentration of 25μL administered by gavage was approximately 4×10 8 CFU Bacillus sp. SLMG-002 and a 25 μL concentration are approximately 4 × 10 8 CFU Bacillus sp. The mice were given a mixed bacterial solution of SLMG-003 once daily for 7 days. The control group continued to consume normal drinking water and feed. During this period, the mice were weighed daily, and their condition, anal sphincter, and diarrhea were observed regularly.

[0030] Conclusion: The body weight of mice in the control group showed a steady increasing trend, while the body weight of mice in the DSS model group decreased from the second day after modeling. Figure 1 During gavage, mouse mortality statistics showed that 3 out of 5 mice in the self-healing group died due to severe diarrhea, with a mortality rate of 60%; only 1 out of 5 mice in the probiotic BS2 group died, with a mortality rate of 20%; and none of the 5 mice in the probiotic BS2+BS3 group died (Table 1). Intestinal length statistics showed that the average intestinal length of mice in the control group was 46.96 cm, while the average intestinal length of mice in the self-healing group shortened to 40.33 cm; the average intestinal length of mice in the probiotic BS2 group was 45.83 cm; and the average intestinal length of mice in the probiotic BS2+BS3 group was 47.55 cm, recovering to the control group level. Figure 2 , Figure 3 ).

[0031] Table 1. Statistical results of mouse mortality during gavage.

[0032] Example 7 Detection of mouse intestinal morphology Mice were euthanized after the gavage experiment, and ileum and colon tissues were collected. The ileum and colon were subjected to HE staining to observe villus length, inflammatory cell infiltration, etc., using the following steps: xylene I and xylene II 10 min each → anhydrous ethanol I 5 min, anhydrous ethanol II 5 min each → 95% ethanol I and 95% ethanol II 3 min each → 80% ethanol 2 min, 70% ethanol and 50% ethanol 2 min each → distilled water 2 min → hematoxylin 2 min → tap water wash 5 min → distilled water 2 min → 1% hydrochloric acid alcohol 3-5 s → 45℃ tap water bluening 5-15 min → distilled water 2 min → 50% ethanol 2 min, 70% ethanol 2 min, 80% ethanol 2 min each → eosin 5 s → 95% ethanol I 2-5 min → 95% ethanol II 2-5 min → anhydrous ethanol I 5 min → anhydrous ethanol II 5 min → xylene I 5 min → xylene II 5-10 min. After staining, resin was applied to the specimens for mounting and air drying. Observations and statistics were then performed.

[0033] Conclusion: In the control group, the ileal villi were densely packed with normal villi length, the colonic crypts were neatly arranged, and there was no inflammatory cell infiltration. In the self-healing group, the ileal villi were sparsely packed with shorter villi, and there was inflammatory cell infiltration in the colon. In the probiotic BS2 group, the ileal villi were denser than those in the self-healing group, the villi length was increased, the colonic crypt structure was more regular, and there was no obvious inflammatory cell infiltration. In the probiotic BS2+BS3 group, the ileal villi were very densely packed with increased villi length, and the colonic crypt structure was more regular. From the perspective of ileal and colonic structure, it was even better than the control group. Figure 4 , Figure 5 ).

[0034] This invention successfully utilizes Bacillus. Bacillus sp. SLMG-002 alleviated DSS-induced colitis in mice, and this method can also be applied in practical applications, using Bacillus subtilis. Bacillus sp. SLMG-002, or Bacillus Bacillus sp. SLMG-002 and Bacillus Bacillus sp. When used in combination with SLMG-003, it regulates the balance of intestinal flora, protects the intestinal mucosal barrier, and inhibits inflammatory responses in the intestine, thereby reducing the threat of colitis to humans and livestock.

[0035] Example 8 Experiment on DSS modeling and DAI index of weaned piglets Thirty weaned piglets were randomly divided into a control group (10 piglets, normally fed) and a DSS treatment group (20 piglets, 0.36g DSS / piglet / day) administered by gavage for 10 days to induce diarrhea. The DSS group was then randomly divided into three subgroups: a self-healing group (no intervention), a probiotic BS2 group (10g DSS / piglet / day administered by gavage daily), and a control group. 9CFU's Bacillus sp. SLMG-002, once daily; probiotics BS2+BS3 group (0.5 x 10 gavage daily). 9 CFU of Bacillus sp. SLMG-002+0.5✖10 9 Bacillus sp. SLMG-003 (CFU) was administered via gavage for 10 consecutive days. The DAI index was calculated and is shown in Tables 2, 3, and 4. After the treatment, some piglets were euthanized, and blood samples were collected for routine blood tests. Serum samples were used to detect immune-related factors, and intestines were collected for paraffin sectioning and HE staining.

[0036] The Diarrhea Index (DAI) assessment procedure was as follows: During the experiment, the defecation status of animals was observed and recorded after DSS gavage and after probiotic treatment, and scored according to the following criteria: normal hard stool = 0 points; soft stool = 1 point; formed moderately loose stool = 2 points; very loose and foamy diarrhea stool = 3 points. The DAI index and its changing trend of each group of animals were calculated to analyze the establishment of the DSS-induced colitis model and the effect of probiotic intervention on the degree of diarrhea.

[0037] in conclusion: Following DSS induction, all weaned piglets developed diarrhea, with 50% or more experiencing severe diarrhea (scoring 3). After probiotic treatment, in the self-healing group, 10% experienced worsening diarrhea, 80% showed no improvement, and 10% experienced relief, indicating that the proportion of piglet diarrhea that resolves or even resolves spontaneously in the short term without intervention is extremely low, approximately 10%. After short-term treatment (10 days) with probiotic BS2, 30% fully recovered, 60% experienced relief, and 10% showed no improvement; no cases of worsening were observed. In the probiotic BS2+BS3 group, after short-term treatment (10 days), 50% fully recovered, 50% experienced relief, and no cases of worsening were observed (see Tables 2, 3, and 4). These results show that adding probiotic BS2 for 10 days achieved a 90% relief rate for piglet diarrhea, and the combined use of BS2 and BS3 for 10 days achieved a 100% relief rate, superior to BS2 alone.

[0038] Table 2 shows the statistics and changes in the DAI index of individuals in the self-healing group.

[0039] Table 3 shows the statistics and changes in DAI index before and after individual experiments in the BS2 probiotic group.

[0040] Table 4 shows the statistics and changes in DAI index before and after individual experiments in the probiotic BS2+BS3 group.

[0041] Example 9 Routine blood tests and serum immune-related factor tests in weaned piglets After the experiment, routine blood tests were performed on weaned piglets. Before blood collection, the pigs were fixed and their chests were exposed. The skin around the anterior vena cava was wiped with 75% alcohol swabs. Using a disposable sterile lancet, the lancet was inserted into the anterior vena cava at a 30°-45° angle towards the opposite shoulder joint at the angle between the right side of the sternal manubrium and the left side of the trachea. After blood collection, the lancet was quickly withdrawn, and 5 mL of whole blood was immediately placed into an EDTA anticoagulant tube. The tube was gently inverted 8-10 times to mix, labeled, and stored at 4°C for routine blood tests. 5 mL of whole blood was injected into a coagulation-promoting tube. The tube was allowed to stand at room temperature until the blood completely coagulated, then centrifuged at 3000 g for 15 minutes. After centrifugation, the centrifuge tube was carefully removed, and the clear serum supernatant was aspirated using a pipette and transferred to a new centrifuge tube, labeled, and numbered. The serum sample was immediately stored at -80°C and transported on dry ice for the detection of immune-related factors.

[0042] Conclusion: White blood cell count showed a significant increase in white blood cell count in the self-healing group compared to the control group, while treatment with probiotic BS2 significantly downregulated white blood cell count, and the white blood cell count in the BS2+BS3 treatment group was at an even lower level. This demonstrates that probiotic BS2 can reduce the level of inflammation in the body, and the combined use of BS2+BS3 can also significantly reduce the level of inflammation in the body. Figure 6 ).

[0043] The results of detection of common inflammatory factors IL-1β, IL-6, IL-8 and TNFα showed that, compared with the control group, the self-healing group of piglets exhibited higher inflammation levels; while after three weeks of treatment with probiotic BS2, their inflammation levels were significantly reduced; the combined use of probiotics BS2 and BS3 had a greater advantage in reducing inflammatory factors, restoring them to levels comparable to or even slightly lower than the control group. Figure 7 ).

[0044] Example 10 Intestinal histological examination of weaned piglets After the experiment, some of the experimental pigs were euthanized, and their intestines were collected. Two-cm tissue blocks were cut from the ileum and colon, placed in centrifuge tubes, fixed in 4% paraformaldehyde for 24 hours, and then dehydrated. The dehydration steps were as follows: 70% alcohol for 2 hours → 80% alcohol for 2 hours → 90% alcohol for 2 hours → 95% alcohol I for 1 hour → 95% alcohol II for 1 hour → 100% alcohol I for 1 hour → 100% alcohol II for 1 hour → Alcohol: xylene (1:1) for 30 minutes → Xylene I for 30 minutes → Xylene II for 30 minutes → Wax soaking for 2 hours → Replace with new wax and soak overnight.

[0045] The next day, the specimens were embedded. After the paraffin solidified, the ileum and colon were stained with hematoxylin and eosin (HE) using the following steps: xylene I and xylene II, 10 min each → anhydrous ethanol I, 5 min each → anhydrous ethanol II, 5 min each → 95% ethanol I and 95% ethanol II, 3 min each → 80% ethanol, 70% ethanol, and 50% ethanol, 2 min each → distilled water, 2 min → hematoxylin, 2 min → tap water wash, 5 min → distilled water, 2 min → 1% hydrochloric acid alcohol, 3-5 s → blueing with tap water at 45℃, 5-15 min → distilled water, 2 min → 50% ethanol, 70% ethanol, and 80% ethanol, 2 min each → eosin, 5 s → 95% ethanol I, 2-5 min → 95% ethanol II, 2-5 min → anhydrous ethanol I, 5 min → anhydrous ethanol II, 5 min → xylene I, 5 min → xylene II, 5-10 min. After staining, resin slides were added to the specimens for mounting and air drying. Observation and statistics were then performed.

[0046] Conclusion: In the control group, the ileal villi were densely packed with normal villi length, the colonic crypts were neatly arranged, and there was no inflammatory cell infiltration. In the self-healing group, the ileal villi were sparsely packed with shorter villi, and there was inflammatory cell infiltration in the colon. In the probiotic BS2 group, the ileal villi were densely packed compared to the self-healing group, the villi length was increased, the colonic crypt structure was more neat, and there was no obvious inflammatory cell infiltration. After the combined use of probiotics BS2 and BS3, the ileal villi were very densely packed, and the villi length even exceeded that of the control group. The colonic crypt structure was neat, and there was no obvious inflammatory cell infiltration. Figure 8 , Figure 9 ).

[0047] This invention successfully alleviated DSS-induced colitis in weaned piglets using Bacillus SLMG-002 strain. The combined use of Bacillus SLMG-002 and Bacillus SLMG-003 showed a synergistic effect, superior to Bacillus SLMG-002 alone in alleviating intestinal inflammation. This method can also be applied in practical applications, using Bacillus SLMG-002 strain or a combination of Bacillus SLMG-002 and SLMG-003 to regulate the balance of intestinal flora, protect the intestinal mucosal barrier, and inhibit inflammatory responses in the intestine.

Claims

1. A strain of Bacillus ( Bacillus sp. ), characterized by: Used to quickly relieve colitis, it is named: Bacillus SLMG-002 strain, and was deposited at the China Center for Type Culture Collection, Wuhan University Collection Center on September 16, 2025, with accession number: CCTCC M 20252231.

2. The application of a Bacillus strain SLMG-002, characterized in that: Bacillus SLMG-002 strain works synergistically through multiple pathways by regulating the balance of intestinal flora, repairing the intestinal mucosal barrier, and inhibiting intestinal inflammatory response, and can be used to prepare a bacterial agent that can quickly relieve colitis.

3. A Bacillus composition, characterized in that: It includes the Bacillus SLMG-002 strain and Bacillus SLMG-003 strain as described in claim 1, in a ratio of 3:7 to 7:

3.

4. The application of a Bacillus composition, characterized in that: The composition of claim 3 works synergistically through multiple pathways by regulating the balance of intestinal flora, repairing the intestinal mucosal barrier, and inhibiting intestinal inflammatory response, and is used to prepare a bacterial agent that rapidly relieves colitis.

5. The application according to any one of claims 2 or 4, characterized in that: The bacterial agents include: liquid bacterial agents and freeze-dried bacterial agents.

6. The application according to claim 5, characterized in that: The liquid bacterial agent is prepared by inoculating Bacillus SLMG-002 or Bacillus SLMG-003 strains into LB liquid medium and culturing them at 37°C in a shaker until the concentration reaches 4 × 10⁻⁶. 8 CFU / 50μL, packaged and refrigerated.

7. The application according to claim 5, characterized in that: The lyophilized bacterial agent was prepared by inoculating Bacillus SLMG-002 or Bacillus SLMG-003 strains into LB liquid medium and culturing them at 37°C in a shaker until the concentration reached 4 × 10⁻⁶. 8 CFU / 50μL, freeze-dried, packaged, and refrigerated.

8. A method for applying a Bacillus inoculant, characterized in that: The Bacillus SLMG-002 inoculant described in claim 2 is added to feed or drinking water at a dosage of 5 × 10⁻⁶. 8 CFU-5×10 9 CFU / kg.

9. A method of applying a Bacillus composition, characterized in that: The Bacillus composition inoculant of claim 4 is added to feed or drinking water at a dosage of 5 × 10⁻⁶. 8 CFU-5×10 9 CFU / kg.

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