Bacillus safensis and application thereof
By using the fermentation broth and feed additives of Bacillus salsa ZHT-9, the research deficiencies of Bacillus salsa in weight loss and fat reduction were addressed, and significant reductions in blood triglycerides and total cholesterol were achieved, along with the regulation of feed intake and body weight.
Patent Information
- Application Number
- CN202511208295.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-08-27
- Publication Date
- 2025-11-21
AI Technical Summary
In the existing technology, there are few studies on Bacillus safranin in terms of weight loss and fat reduction, and its role in animal weight regulation and lipid metabolism has not been clearly explored.
A strain of Bacillus salsa ZHT-9 was provided, which, through the preparation of fermentation broth and feed additives, reduced the levels of triglycerides and total cholesterol in the blood of animals and regulated the relationship between feed intake and weight gain.
Bacillus salsa ZHT-9 significantly reduced blood triglyceride and total cholesterol levels in mice, pigs, and ruminants, and regulated their food intake and weight gain, exhibiting a "eat more but not gain weight" phenomenon, filling a research gap in the lipid-lowering function of this strain.
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Abstract
Description
Technical Field
[0001] This invention belongs to the field of microbiology, specifically relating to a strain of Bacillus safranin and its applications. Background Technology
[0002] Currently, probiotics used for weight loss and fat reduction mainly include Lactobacillus (such as Lactobacillus gasseri, Lactobacillus reuteri, Lactobacillus plantarum, Lactobacillus rhamnosus, Bifidobacterium lactis, and Bifidobacterium longum) and Bifidobacterium (such as Bifidobacterium animalis subsp. lactis B420, Bifidobacterium breve B-3, Lactobacillus gasseri, and Lactobacillus paracasei). These probiotics have significant effects on weight loss by regulating gut microbiota, improving energy metabolism, inhibiting fat accumulation, reducing inflammatory markers, increasing satiety, and increasing the production of short-chain fatty acids. However, there is currently limited research on Bacillus in weight loss and fat reduction, and existing literature does not explicitly mention the specific role of Bacillus saforiti in fat reduction.
[0003] Bacillus sarfusae ( Bacillus safensis It is widely used in agriculture, mainly in the following aspects: 1. Promotes plant growth: By secreting plant hormones such as indoleacetic acid (IAA), it stimulates root development, enhances photosynthesis, and increases crop yield. At the same time, it decomposes soil organic matter, releasing nutrients such as nitrogen, phosphorus, and potassium, which are then absorbed by plants.
[0004] 2. Biological control: It can inhibit pathogens such as tomato gray mold and tobacco black shank by producing antibiotics and antimicrobial proteins to directly suppress pathogenic microorganisms. It can also control root-knot nematodes, reducing the use of chemical pesticides.
[0005] 3. Improve soil and nutrient cycle: decompose organic matter to improve soil fertility; secrete organic acids and enzymes to activate insoluble phosphorus and potassium, improve nutrient utilization, and reduce dependence on chemical fertilizers.
[0006] 4. Strong resistance to adverse conditions: It can withstand drought, salinity and other adverse conditions. Its spore structure is resistant to high temperature and drying, making it easy to store and transport, and adaptable to a variety of agricultural environments.
[0007] 5. Collaborative application in industry and agriculture: The cellulase, protease, and other enzymes produced can be used in industry (such as papermaking and biodegradation) while promoting sustainable agricultural development.
[0008] 6. Ecological balance: As beneficial microorganisms, they maintain the balance of soil microbial communities, reduce the reproduction of pathogens, reduce pesticide residues, and contribute to green agriculture.
[0009] There are currently no research reports on Bacillus safortiformis (Sarfus). Bacillus safensisIt has the ability to resist acid and choline salt and the effect of reducing animal weight or regulating blood lipid. The main research and application of the bacteria is still concentrated in the field of agriculture, such as promoting plant growth, biological control, soil improvement, etc. If future research explores its potential in animal health or metabolic regulation, it may further expand its application range. SUMMARY
[0010] To solve the above problems, the present application provides a strain of Bacillus safensis, which can effectively reduce the triglyceride (TG) and total cholesterol (TC) levels in the blood, thereby improving blood lipid metabolism, and can significantly regulate the relationship between food intake and body weight gain in mice, pigs and ruminants.
[0011] To solve the problems existing in the prior art, the technical scheme adopted by the present application is: In a first aspect, the present application provides a strain of Bacillus safensis ZHT-9, the preservation number of which is CCTCC NO: M20241011; the preservation date is May 20, 2024; and the preservation unit is China Center for Type Culture Collection.
[0012] In a second aspect, the present application provides the use of the Bacillus safensis ZHT-9 of the first aspect in the preparation of a product for improving animal growth performance, reducing blood lipid and regulating body weight.
[0013] Further, the blood lipid reduction refers to reducing the triglyceride (TG) and total cholesterol (TC) levels in the blood.
[0014] Further, the animal is a mouse, a pig or a ruminant.
[0015] In a third aspect, the present application provides a product, which comprises the Bacillus safensis ZHT-9 of the first aspect, a fermentation broth of the Bacillus safensis ZHT-9, a fermentation broth supernatant, a fermentation broth precipitate, live bacteria and / or dead bacteria.
[0016] Further, the product is a feed additive.
[0017] Further, the number of live Bacillus safensis ZHT-9 in the feed additive is not less than 5 x 10 7 cfu / g.
[0018] Further, the number of live Bacillus safensis ZHT-9 in the feed additive is 1 x 10 9 cfu / g.
[0019] Further, the fermentation broth refers to a liquid obtained by inoculating a strain into a culture medium and culturing for a period of time.
[0020] Furthermore, the supernatant of the fermentation broth refers to the clear liquid at the top after centrifugation of the fermentation broth; it contains abundant metabolic products from the bacterial growth and reproduction process, as well as some bacterial cell fragments. The acidic substances and bacteriocins secreted by the bacteria have antagonistic and killing effects on harmful bacteria. The amino acids and vitamins synthesized by the bacteria after decomposing food are also in the culture medium, as well as enzymes secreted by the bacteria that are useful to the human body. Some of the bacterial cell components also have an immune-boosting effect on the human body.
[0021] Furthermore, the fermentation broth precipitation refers to the liquid precipitate obtained by centrifugation, which includes free protein, residual bacterial cells, broken cells, and culture medium residue, mainly protein and intracellular matrix.
[0022] Furthermore, the live bacteria mentioned refer to active bacterial flora that can colonize and multiply in the intestines, which helps increase the number of beneficial bacteria.
[0023] Furthermore, the term "dead bacteria" refers to microorganisms that have lost their vitality and are unable to grow and reproduce. Probiotics lose their vitality due to the production process, such as high-temperature treatment or excessive drying.
[0024] Fourthly, the present invention provides a method for preparing the fermentation broth of Bacillus sarcodactylis ZHT-9 as described in the third aspect above, comprising the following steps: Bacillus sabolicus ZHT-9 was inoculated onto TSA medium and cultured at 37±1℃ for 24±1h to activate the strain. Single colonies were picked and inoculated into TSB medium and cultured with shaking at 37±1℃ and 180±5rpm for 16±1h. The next day, fresh bacterial culture was inoculated into TSB medium and fermented at 37±1℃ and 180±5rpm for 16±1h to obtain the fermentation broth.
[0025] Furthermore, the fresh bacterial culture was inoculated into TSB medium at an inoculation rate of (1±0.1)%v / v.
[0026] Preferably, the concentration of viable bacteria reaching the bacterial growth plateau in the fermentation broth is (7.53±4.38) x 10⁻⁶. 7 cfu / ml.
[0027] The advantages and beneficial effects of this invention are: Bacillus sarfusae ( Bacillus safensis ZHT-9 exhibits significant lipid-lowering functions in the blood of mice, pigs, and ruminants. Animal experiments have confirmed that this strain can effectively reduce blood triglyceride (TG) and total cholesterol (TC) levels, thereby improving lipid metabolism. This discovery fills a gap in the research on the lipid-lowering function of Bacillus safranin and provides a novel microbial resource for the prevention and treatment of hyperlipidemia and related metabolic diseases.
[0028] The Bacillus safensis ZHT-9 has a wide adaptation range. Animal feeding experiment results show that, although the food intake of mice, pigs and ruminants ingesting the ZHT-9 strain increases, the body weight growth of the mice, pigs and ruminants is significantly lower than that of the control group, indicating that the Bacillus safensis ZHT-9 can significantly regulate the relationship between food intake and body weight growth of mice, pigs and ruminants.
[0029] The "eat more and not gain weight" phenomenon exhibited by the Bacillus safensis ZHT-9 is a unique physiological effect that distinguishes it from other known strains. This effect is not only effective for monogastric animals, but also can regulate the healthy metabolism of ruminants.
[0030] Preservation instructions: Strain name: Bacillus safensis ZHT-9 Preservation number: CCTCC NO: M20241011 Classification name: Bacillus safensis Preservation date: May 20, 2024 Preservation unit: China Center for Type Culture Collection Address of the preservation unit: Wuhan University, Wuhan, China. Brief description of the drawings
[0031] Figure 1 The figure is the colony morphology of the Bacillus safensis ZHT-9; Wherein: a is the colony plate morphology of ZHT-9; b is the colony morphology of ZHT-9 under gram staining microscopic examination (oil lens 100x, ocular lens 10x); Figure 2 The figure is the growth curve of the Bacillus safensis ZHT-9; Figure 3 The figure is a phylogenetic tree of the Bacillus safensis ZHT-9 constructed based on pan-genome using Neighbou-joining clustering. DETAILED DESCRIPTION
[0032] The technical solutions in the embodiments of the present application will be clearly described below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art belong to the scope of protection of the present application.
[0033] In the following examples, the experimental methods are conventional methods, and are performed according to the techniques or conditions described in the literature in the art or according to the product instructions, unless otherwise specified. The materials, reagents, etc. used in the following examples can be obtained from commercial channels, unless otherwise specified.
[0034] The Bacillus safensis ZHT-9 of the present application is referred to as ZHT-9.
[0035] Strain source: The Bacillus safensis ZHT-9 used for isolation and screening in the present application is from a pig farm in Qionglai City, Sichuan Province.
[0036] TSB medium (OXOID, UK) and TSA medium (OXOID, UK) are used for culturing the bacteria ZHT-9.
[0037] SPF mice are 4 weeks old and purchased from Chengdu Dashuo Experimental Animal Co., Ltd.
[0038] The Chenghua black pig fattening pigs used in the test are from a pig farm in Pengzhou City, Sichuan Province.
[0039] The ruminant (ma sheep) used in the test is from a Chengdu ma sheep farm in Dayi County, Sichuan Province.
[0040] Taq DNA polymerase, dNTP, and bacterial genomic DNA extraction kit are purchased from Takara Bio Co., Ltd.
[0041] The triglyceride (TG) detection kit is purchased from Beijing Qisong Biological Co., Ltd.
[0042] The blood index detector is Seamaty-120VP, which can be used to detect various biochemical indexes in blood.
[0043] The Bacillus safensis ZHT-9 of the present application is referred to as ZHT-9.
[0044] Strain source: The Bacillus safensis ZHT-9 used for isolation and screening in the present application is from a pig farm in Qionglai City, Sichuan Province.
[0045] Example 1
[0046] 1. Isolation, purification and identification of Bacillus safensis ZHT-9 1.1 Isolation, purification, staining and microscopic examination of the strain The intestinal contents of Chenghua black pigs are spread on TSA plates, and single colonies are picked and purified by plate streaking. Single colonies with good growth are picked from the bacterial purified culture medium to obtain strain ZHT-9, which is subjected to Gram staining. The morphological characteristics of the bacterial body are observed by light microscopy.
[0047] As shown in Figure 1 a, the strain grows on TSA medium for 24h, and the colony diameter is 2-5mm, the colony is milky white, the edge is smooth, and the center is wrinkled.
[0048] 1.2 Physiological and biochemical identification: AsFigure 1 b, the bacteria is gram-positive (G+), and the cell shape is long rod, with a cell length of 1-1.5 pm and a width of 0.3-0.5 pm.
[0049] 1.3 Determination of growth curve Fresh single colonies on TSA medium (OXOID, UK) were picked and inoculated into a bacterial flask containing TSB medium (OXOID, UK), which was cultured in a 37 °C shaker at a speed of 180 rpm / min. Every 2 h, the bacteria were taken out and the OD 630 Bacterial growth curve was drawn. After 12 hours of culture in TSB medium, the bacteria entered the plateau phase of growth Figure 2 ).
[0050] 1.4 Molecular identification and preservation of bacteria The purified strain ZHT-9 was used to extract the genomic DNA of the strain ZHT-9 using a bacterial genomic extraction kit (Takara Company). The extracted DNA was subjected to 16S rRNA gene PCR amplification and sequencing splicing by a sequencing company. The bacterial 16S rDNA was amplified by primers 27F / 1492R, wherein the upstream primer 27F (SEQ ID NO. 1: 5'-AGAGTTTGATCCTGGCTCAG-3') and the downstream primer 1492R (SEQ ID NO. 2: 5'-GGTTACCTTGTTACGACTT-3'). The PCR reaction system (50 pl) was as follows: 5x PS Buffer 10 pl, 10 pmol / L 27F 5 pl; 10 pmol / L 1492R 5 pl; 2.5 pmol / L dNTP 4 pl; bacterial liquid template 4.5 pl; Primer STAR 1.5 pl; H2O 20 pl. The PCR reaction conditions were as follows: 98 °C for 5 min; 98 °C for 10 sec, 55 °C for 5 sec, 72 °C for 60 sec, and this step was repeated for 30 cycles; 72 °C for 10 min. The strain was sent to Shengong Bioengineering Co., Ltd. for whole genome sequencing.
[0051] Based on the pan gene, the phylogenetic tree of the bacteria was constructed by Neighbou-joining clustering, as shown in Figure 3 .
[0052] 1.5 Identification results: The 16S rRNA gene fragment of the amplified bacteria has a size of 1456 bp, and the 16S rRNA sequence obtained by sequencing is shown as SEQ ID NO. 3, and compared with (bacteria in NCBI database), the results show that the bacteria belongs to Bacillus, and through whole genome sequencing, the strain ZHT-9 is identified as Bacillus safensis. The genome accession information of the bacteria is JAZDWY010000000, and the species name is Bacillus safensis .
[0053] The Bacillus safensis ZHT-9 is preserved in the China Center for Type Culture Collection, the preservation number is CCTCC NO: M20241011, the preservation address is Wuhan University in Wuhan, China, and the preservation date is May 20, 2024.
[0054] SEQ ID NO. 3: 2. Sub-culturing of the bacteria
[0055] Bacillus safensis ZHT-9 was inoculated on TSA medium (OXOID, UK) and incubated at 37 °C for 24 h to activate the strain. A single colony was picked and inoculated in TSB medium and incubated at 37 °C, 180 rpm for 16 h. The next day, fresh bacterial solution was inoculated in TSB medium (OXOID, UK) at 1% v / v and incubated at 37 °C, 180 rpm for 16 h to obtain the fermentation broth. The viable cell concentration of the bacteria at the stationary phase reached (7.53 ± 4.38) x 10 7 cfu / ml.
[0056] 3. In vitro resistance screening of Bacillus safensis ZHT-9.
[0057] 3.1 Acid resistance test TSB medium (OXOID, UK) was adjusted to pH 2.0 and 4.0 respectively and sterilized at 121 °C for 15 min. 100 ml of the fermentation broth of the bacteria at the logarithmic phase was inoculated in 900 ml of the TSB medium with the corresponding pH and incubated for 3 h. Then, the bacteria were diluted by 2-fold and inoculated on TSA solid medium and incubated at 37 °C for 24 h. The next day, the survival number of the strain was determined by plate counting.
[0058] 3.2 Cholate salt resistance test Pig cholate salt TSB medium was prepared at concentrations of 0.2% and 0.5% and filtered to a sterilized flask with a 0.22 μm filter, and TSB medium without pig cholate salt was used as a negative control. 100 ml of the fermentation broth of the bacteria at the logarithmic phase was inoculated in 900 ml of the TSB medium with pig cholate salt at the corresponding concentration and incubated for 2 h. Then, the bacteria were diluted by 2-fold and inoculated on TSA medium and incubated at 37 °C for 24 h. The next day, the survival number of the strain was determined by plate counting.
[0059] Table 1 Influence of low pH on the survival rate of strain ZHT-9
[0060] As shown in Table 1, the survival rate of the strain of the application in the medium with pH 2.0 reached 52.94% after 3 h of incubation, and the survival rate of the strain of the application in the medium with pH 4.0 reached 88.24% after 3 h of incubation, indicating that the strain of the application has certain acid resistance.
[0061] Table 2 Influence of pig cholate salt at concentrations of 0.2% and 0.5% on the survival rate of strain ZHT-9
[0062] As shown in Table 2, the survival rate of the strain of the application can reach 12.3% after 2h of culture in a medium with a 0.2% concentration of bile salt; the survival rate of the strain of the application can reach 1.22% after 2h of culture in a medium with a 0.5% concentration of bile salt, indicating that the strain of the application has a certain bile salt tolerance.
[0063] Experimental Example 1 Animal feeding test 1. Experimental method The experimental animals were 36 SPF female Kunming mice aged 4 weeks, which were randomly divided into 3 groups, 12 in each group, namely a control group, a high-dose test group (ZHT-9G) and a low-dose test group (ZHT-9D).
[0064] The control group was fed with conventional SPF feed, and the test groups (ZHT-9) were all fed with conventional SPF feed (purchased from Chengdu Dashuo Experimental Animal Co., Ltd., nutritional components as follows: moisture ≤10%, crude protein ≥18%, crude fat ≥4%, crude fiber ≤5%, crude ash ≤8%, 1.0%≤calcium≤1.8%, 0.6%≤phosphorus≤1.2%, lysine ≥0.82%, methionine + cystine ≥0.53%) on the basis of adding Bacillus safensis ZHT-9, and the mice were free to eat. The high-dose test group (ZHT-9G) reached 5 x 10 8 cfu / g of feed, and the low-dose test group (ZHT-9D) reached 5 x 10 7 cfu / g of feed, and the bacterial content of the high-dose group was 10 times that of the low-dose group. The pre-test period of the test was 7 days, and the formal period was 30 days. During the experiment, the body weight and feed intake of the mice were recorded every day. After the test, the total cholesterol blood index of the mice was detected by eyeball blood collection (with heparin sodium anticoagulant). The whole blood without anticoagulant was centrifuged at 3000r / min x 3min after standing at 4℃ overnight, and the serum sample of the mice was collected, and the cholesterol and triglyceride indexes were detected by using the kit.
[0065] 2. Results and analysis 2.1 Animal serum index From Table 3, after 30 days of formal feeding test, the triglyceride in blood of mice in ZHT-9G and ZHT-9D groups was 3422.25 ± 129.85 nmol / L and 3486 ± 114.83 nmol / L respectively, compared with 3537.25 ± 191.89 nmol / L of the control group, both of which were significantly decreased, and the decrease amplitude was positively correlated with the ZHT-9 dose. The total cholesterol in blood of mice in ZHT-9G and ZHT-9D groups was 1.39 ± 0.17 mmol / L and 1.57 ± 0.14 mmol / L respectively, compared with 1.73 ± 0.12 mmol / L of the control group, both of which were significantly decreased, and the decrease amplitude was positively correlated with the ZHT-9 dose.
[0066] Table 3 Effect of ZHT-9 on blood lipid level of mice
[0067] Note: * means that the number has significance (P≤0.05) compared with the control group.
[0068] 2.2 Animal growth From Table 4, the initial weight of the control group, ZHT-9G and ZHT-9D groups had no obvious difference. After 30 days of formal feeding test, the final weight, average daily feed intake and average daily gain of mice in ZHT-9G group were 45.69 ± 1.31 g, 5.8 ± 1.24 g and 0.61 ± 0.38 g respectively; the final weight, average daily feed intake and average daily gain of mice in ZHT-9D group were 46.08 ± 1.42 g, 5.53 ± 1.08 g and 0.62 ± 0.56 g respectively, all of which had significant difference with the control group, and the weight loss amplitude was positively correlated with the bacterial dose, and the feed intake was negatively correlated with the bacterial dose. The results showed that the mice appeared "eat more and not fat" phenomenon.
[0069] Table 4 Effect of ZHT-9 on growth performance of mice
[0070] Note: * means that the number has significance (P≤0.05) compared with the control group.
[0071] Experimental Example 2 Fattening pig feeding test
[0072] 1. Experimental method Experimental animals were selected 35 kg or so of Chenghua black pig fattening pigs 20, were randomly divided into 2 groups, each group of 10, a group as control group, a group as test group (ZHT-9). The control group was fed with conventional feed (purchased from the whole Wei feed technology Co., Ltd., the main raw materials are corn, soybean meal, fish meal, mineral elements, compound vitamins and appropriate fungicides, the nutritional composition is as follows: moisture ≤14%, crude fiber ≤5%, crude ash ≤8%, 0.5%≤ calcium ≤1.0%, phosphorus ≥0.5%, lysine ≥1.102%, 0.3%≤ calcium ≤0.8%, the test group (ZHT-9) was added with bacillus safensis ZHT-9 on the basis of feeding conventional feed, so that the number of viable bacteria per gram of feed reached 1 x 10 9 cfu. The test pretest period was 7 days, and the formal period was 30 days. During the experiment, the body weight and feed intake of the fattening pigs were recorded. After the experiment, the whole blood (with anticoagulant) of the fattening pigs was collected and the triglyceride and cholesterol indexes were detected by Seamaty-120VP detector.
[0073] 2 Results 2.1 Animal serum indexes
[0074] From table 5, after 37 days of test feeding, the triglyceride and total cholesterol in the blood of Chenghua black pigs in ZHT-9 group were 0.57 ± 0.05 mmol / L and 2.18 ± 0.14 mmol / L, respectively, which were significantly lower than those in the control group, 0.79 ± 0.07 mmol / L and 2.53 ± 0.16 mmol / L, respectively. The results showed that the bacteria ZHT-9 had significant hypolipidemic function in the blood of live pigs.
[0075] Table 5 Effect of ZHT-9 on blood lipid level of fattening pigs
[0076] Note: * indicates that compared with the control group, the number has significant (P≤0.05).
[0077] 2.2 Animal growth From table 6, there was no significant difference in initial weight and average daily feed intake between the control group and ZHT-9 group. The final weight, average daily feed intake, average daily gain and feed conversion ratio of ZHT-9 group were 48.85 ± 6.35 kg, 2.11 ± 0.05 kg / d, 0.35 ± 0.06 kg / d and 6.02 ± 0.83, respectively, which were significantly different from those of the control group. The results showed that the live pig appeared "eat more and not fat" phenomenon.
[0078] Table 6 Effect of ZHT-9 on growth performance of Chenghua black pigs
[0079] Note: * means that the number is significant (P < 0.05) compared with the control group.
[0080] Experiment Example 3 Ruminant feeding test 1. Experimental method The experimental animals were 30 fattening sheep of about 4-50 kg, which were randomly divided into two groups, each group of 15. One group was the control group, and the other group was the test group (ZHT-9). The control group was fed with conventional feed (corn silage feed), and the test group (ZHT-9) was added with Bacillus safensis ZHT-9 on the basis of feeding conventional feed, so that the number of viable bacteria per gram of feed reached 1 x 10 9 cfu. The pre-test period of the test was 4 days, and the formal period was 24 days. During the experiment, the body weight and feed intake of the sheep were recorded. After the end of the test, the whole blood of the sheep (with anticoagulant) was collected for detection of triglyceride and cholesterol indexes by Seamaty-120VP detector.
[0081] 2. Results 2.3 Serum indexes of animals As shown in Table 7, after feeding for 28 days, the triglyceride and total cholesterol in the blood of the sheep in the ZHT-9 group were 0.43 ± 0.04 mmol / L and 1.87 ± 0.56 mmol / L, respectively, which were significantly lower than 0.48 ± 0.05 mmol / L and 2.06 ± 0.32 mmol / L of the control group. The results showed that the bacteria ZHT-9 had a significant blood lipid-lowering function in the ruminant body.
[0082] Table 7 Effect of ZHT-9 on blood lipid level of sheep
[0083] Note: * means that the number is significant (P < 0.05) compared with the control group.
[0084] 2.4 Growth of animals As shown in Table 8, there was no significant difference in initial weight, final weight and average daily feed intake between the control group and the ZHT-9 group. The average daily gain and the feed conversion ratio of the sheep in the ZHT-9 group were 0.09 ± 0.03 kg / d and 15.10 ± 4.54, respectively, which were significantly different from those of the control group. The results showed that the ruminant showed the phenomenon of "eating more but not gaining weight".
[0085] Table 8 Effect of ZHT-9 on growth performance of sheep
[0086] Note: * means that the number is significant (P < 0.05) compared with the control group.
[0087] From the experimental examples 1-3, it can be seen that the body weight growth of the mice, pigs and ruminants ingesting the ZHT-9 strain is significantly lower than that of the control group, although the food intake is increased, which indicates that the Bacillus safensis ZHT-9 can significantly regulate the relationship between the food intake and the body weight growth of the mice, pigs and ruminants. The "eat more and not gain weight" effect of the Bacillus safensis ZHT-9 is not only effective for monogastric animals, but also can regulate the healthy metabolism of ruminants, and has the possibility of being applied to humans.
[0088] The embodiments of the present application are described above in combination with the drawings, but the present application is not limited to the specific embodiments described above, and the specific embodiments described above are only illustrative but not restrictive, and those of ordinary skill in the art can make many forms under the inspiration of the present application without departing from the scope of the present application and the scope protected by the claims, which all belong to the protection of the present application.
Claims
1. A strain of Bacillus sarcodactylis ZHT-9, characterized in that, The accession number of the *Bacillus sarcodactylis* ZHT-9 is CCTCC NO: M20241011; accession date: May 20, 2024. Depository: China Center for Type Culture Collection.
2. The use of Bacillus sarcodactylis ZHT-9 as described in claim 1 in the preparation of products that improve animal growth performance, lower blood lipids, and regulate body weight.
3. The application according to claim 2, characterized in that, The animal in question is a rat, a pig, or a ruminant.
4. A product characterized in that, Includes Bacillus salsa ZHT-9 as described in claim 1, fermentation broth of Bacillus salsa ZHT-9, fermentation broth supernatant, fermentation broth precipitate, live bacteria and / or dead bacteria.
5. The product according to claim 4, characterized in that, The product is a feed additive.
6. The product according to claim 5, characterized in that, The viable count of Bacillus salsa ZHT-9 in the feed additive is not less than 5 x 10⁻⁶. 7 cfu / g.
7. The product according to claim 6, characterized in that, The viable count of Bacillus salsa ZHT-9 in the feed additive is 1 x 10⁻⁶. 9 cfu / g.
8. The method for preparing the fermentation broth of Bacillus salsa ZHT-9 according to claim 4, characterized in that, Includes the following steps: Bacillus sabolicus ZHT-9 was inoculated onto TSA medium and cultured at 37±1℃ for 24±1h to activate the strain. Single colonies were picked and inoculated into TSB medium and cultured with shaking at 37±1℃ and 180±5rpm for 16±1h. The next day, fresh bacterial culture was inoculated into TSB medium and fermented at 37±1℃ and 180±5rpm for 16±1h to obtain the fermentation broth.
9. The method for preparing the fermentation broth of Bacillus salsa ZHT-9 according to claim 8, characterized in that, The fresh bacterial culture was inoculated into TSB medium at an inoculation rate of (1±0.1)%v / v.