Micro-ecological powder as well as preparation method and application thereof

Abstract

The invention discloses micro-ecological powder as well as a preparation method and application thereof, and belongs to the technical field of microorganisms. The feed additive comprises compound zymophyte powder and bacillus subtilis powder, the compound zymophyte powder is prepared from lactobacillus plantarum CGMCC (China General Microbiological Culture Collection Center) NO.12849 and enterococcus faecium CGMCC NO.9134; the bacillus subtilis bacterial powder is prepared from bacillus subtilis CGMCC (China General Microbiological Culture Collection Center) NO.25499. The micro-ecological powder has the characteristics of wide antibacterial spectrum, high efficiency, no cytotoxicity, no hemolysis, high stability and the like. The micro-ecological powder provided by the invention has remarkable advantages, can efficiently inhibit gram-negative pathogenic bacteria and gram-positive pathogenic bacteria at the same time, achieves the effect of resisting both yin and yang, can reduce the diarrhea rate of livestock and poultry and promote the growth of livestock and poultry, and has a wide application prospect.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms, and specifically relates to a microecological powder and a preparation method and application thereof. Background Art

[0002] Most of the existing antibacterial products usually do not have a specific target, and their antibacterial effects are often only directed against a certain type of pathogens, such as Gram-positive pathogens or Gram-negative pathogens.

[0003] Probiotics are biologically active preparations made from living microorganisms, which can effectively improve the balance of intestinal flora in livestock and poultry, and are conducive to the healthy growth of livestock and poultry. In addition, probiotics also have the characteristics of no residue, no toxic side effects, etc. Therefore, how to develop a new antibacterial probiotic with a broad spectrum, high efficiency and low toxicity has a very important practical application value. The probiotic powder of the present invention can efficiently inhibit Gram-negative pathogens and Gram-positive pathogens at the same time, achieving the effect of yin and yang dual resistance, and providing a basis for the application of probiotics in livestock and poultry production. Summary of the invention

[0004] In order to solve the above technical problems, the present invention provides a microecological powder, which includes composite fermented bacterial powder and Bacillus subtilis powder; the composite fermented bacterial powder is made of Lactobacillus plantarum CGMCC NO.12849 and Enterococcus faecium CGMCC NO.9134; the Bacillus subtilis powder is made of Bacillus subtilis CGMCC NO.25499.

[0005] Furthermore, the composite fermentation bacteria powder and the Bacillus subtilis bacteria powder are prepared according to (1-10): (1-3).

[0006] Furthermore, the composite fermentation bacteria powder and the Bacillus subtilis bacteria powder are prepared in a ratio of 7:3.

[0007] The present invention also provides the use of the microecological powder in preparing medicine or feed for inhibiting Gram-negative bacteria and / or Gram-negative bacteria.

[0008] Furthermore, the Gram-negative bacteria are Escherichia coli and Salmonella, and the Gram-positive bacteria are Staphylococcus aureus and Clostridium perfringens.

[0009] The present invention also provides a microecological preparation of the microecological powder.

[0010] The invention also provides feed containing the microecological powder.

[0011] The present invention also provides the use of the microecological powder in preparing medicine or feed for improving the growth performance of livestock and poultry.

[0012] The present invention also provides a method for preparing the microecological powder, which is characterized by being prepared according to the following steps:

[0013] (1) Lactobacillus plantarum CGMCC No. 12849 and Enterococcus faecium CGMCC No. 9134 were inoculated into a fermentation medium at a ratio of 1:1 for fermentation. After the fermentation was completed, the supernatant of the fermentation liquid after centrifugation was spray-dried into a powder;

[0014] (2) inoculating Bacillus subtilis CGMCCNO.25499 into a fermentation medium for fermentation, and after the fermentation is completed, filtering the fermentation liquid and spray-drying it into a powder;

[0015] (3) The bacterial powders obtained in step (1) and step (2) are mixed evenly according to a certain proportion to obtain the product.

[0016] Furthermore, after the fermentation is completed, the number of viable bacteria in the fermentation liquid is (1-9)×10 10 CFU / mL.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] The microecological powder provided by the present invention can inhibit both Gram-negative and Gram-positive pathogens, and has the characteristics of broad-spectrum antibacterial, high efficiency, no cytotoxicity, high stability, etc. The microecological powder formula provided by the present invention can significantly reduce the diarrhea rate of livestock and poultry and promote the growth of livestock and poultry. The antibacterial microecological powder is easy to produce on a large scale and has low cost, and has a very broad application prospect. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 The results of the minimum inhibitory concentration (maximum dilution multiple) of different bacterial powders for different pathogenic bacteria are shown in Figure 1, where: 1 is a composite bacterial agent of Lactobacillus plantarum CGMCC NO.12849 and Enterococcus faecium CGMCC NO.9134; 2 is a microecological powder composed of a composite bacterial agent of Lactobacillus plantarum CGMCC NO.12849 and Enterococcus faecium CGMCC NO.9134 plus Bacillus subtilis CGMCC NO.25499 powder; 3 is a microecological powder composed of a composite bacterial agent of Lactobacillus plantarum CGMCC NO.12849 and Enterococcus faecium CGMCC NO.9134 plus Bacillus subtilis CGMCC NO.17348 powder; 4 is a microecological powder composed of a composite bacterial agent of Lactobacillus plantarum CGMCC NO.12849 and Enterococcus faecium CGMCC NO.9134 plus Bacillus veleus CGMCC NO.27887 powder.

[0020] Figure 2 This is the result graph of the minimum inhibitory concentration (maximum dilution multiple) of the optimal formula antibacterial microecological powder against different pathogenic bacteria. DETAILED DESCRIPTION

[0021] The embodiments of the present invention will be described in detail below in conjunction with the examples, but it will be appreciated by those skilled in the art that the following examples are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. If no specific conditions are specified in the examples, the conditions are carried out according to normal conditions or manufacturer recommendations. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be purchased from the market.

[0022] Example 1 Screening of Gram-negative pathogenic bacteria

[0023] The experimental strains of this embodiment are:

[0024] Lactobacillus plantarum CGMCC NO.12849 is a strain owned by our laboratory. The strain is named Lactobacillus plantarum DBNZW02 and was deposited on August 15, 2016 at the General Microbiology Center of China Culture Collection Administration, referred to as CGMCC, at No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, with a deposit number of CGMCC No. 12849. For details, see patent CN108570421B.

[0025] Enterococcus faecium CGMCC NO.9134 is a strain of our laboratory, named Enterococcus faecium UE01, which was deposited on May 8, 2014 at the General Microbiology Center of China Culture Collection Administration, address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, with the deposit number CGMCC No.9134. For details, see patent CN104388327B.

[0026] Lactobacillus reuteri CGMCC NO.19493 is a strain of our laboratory, named Lactobacillus reuteri DBN-JP, which was deposited at the General Microbiology Center of China National Committee for the Preservation of Microorganisms on March 19, 2020, at the Institute of Microbiology, Chinese Academy of Sciences, No. 3, No. 1, Beichen West Road, Chaoyang District, Beijing, with a deposit number of CGMCC No. 19493. For details, see patent CN112011481B.

[0027] Lactobacillus acidophilus CGMCC No. 5093 is a strain owned by our laboratory. The applicant has deposited it in the General Microbiological Center of China National Committee for Microbiological Culture Collection (hereinafter referred to as CGMCC) on July 26, 2011. The address is: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, with the deposit number CGMCC No. 5093. For details, see patent CN102559539A.

[0028] 1. Culture medium preparation:

[0029] MRS liquid culture medium, the composition and content are as follows: yeast powder 5g / L, glucose 20g / L, dipotassium hydrogen phosphate 2g / L, diammonium citrate 2g / L, sodium acetate 3g / L, magnesium sulfate 0.2g / L, manganese sulfate 0.05g / L, Tween 80 1mL / L.

[0030] 2. Primary activation:

[0031] Single colonies of Lactobacillus plantarum CGMCC NO.12849, Enterococcus faecium CGMCC NO.9134, Lactobacillus reuteri CGMCC NO.19493 and Lactobacillus acidophilus CGMCC No.5093 were inoculated into MRS liquid culture medium, and cultured anaerobically at 37°C for 24 hours to obtain primary culture fermentation liquid.

[0032] 3. Antibacterial scheme and effect evaluation:

[0033] The first-level fermentation broth of Lactobacillus plantarum CGMCC NO.12849, Enterococcus faecium CGMCC NO.9134 and Lactobacillus reuteri CGMCC NO.19493 was inoculated into MRS liquid culture medium at an inoculation amount of 1% and cultured at 37°C for 24 hours to obtain the second-level fermentation broth. After the fermentation was completed, the number of viable bacteria in the fermentation broth was (1-3)×10 9 CFU / mL, and the antibacterial evaluation was performed by the Oxford cup method.

[0034] Antibacterial evaluation method:

[0035] The double-layer agar Oxford cup method was used to determine the inhibitory diameter of the samples against Escherichia coli K88 and Salmonella SL1344.

[0036] In the clean bench, pour about 7-10mL of sterilized 0.8% plain agar into each sterilized petri dish to make the bottom layer uniform. After the plain agar cools and solidifies, place the sterilized Oxford cup on the agar. Dilute the indicator bacteria cultured overnight by 200 times and add it to the culture medium at a constant temperature of 50±5℃ at a ratio of 1%. Shake it gently to avoid foaming. Pour the culture medium with bacterial solution into the water agar culture dish with the Oxford cup placed. Pour 20mL into each culture dish. The thickness of the bacterial layer in each culture dish should be uniform.

[0037] After the above culture medium cools and solidifies, use tweezers to remove the Oxford cup and mark it on the culture dish. Pipette 150μL of the sample solution to be tested and add it to the corresponding well. Diffusion at room temperature for about 1 to 2 hours. Transfer the culture dish with the sample to a 37℃ incubator for incubation and observe the results after 12-16 hours of incubation. Use a vernier caliper to measure the diameter of the antibacterial zone in the plate to determine the antibacterial effect of the sample.

[0038] The antibacterial scheme and ratio are as follows:

[0039] (1) Secondary fermentation broth of Lactobacillus plantarum CGMCC NO.12849;

[0040] (2) Lactobacillus acidophilus CGMCC NO.5093 secondary fermentation broth;

[0041] (3) Secondary fermentation broth of Enterococcus faecium CGMCC No.9134;

[0042] (4) Lactobacillus reuteri CGMCC NO.19493 secondary fermentation broth;

[0043] (5) inoculating the primary fermentation broth with Lactobacillus plantarum CGMCC NO.12849 and Lactobacillus acidophilus CGMCC NO.5093 (1:1);

[0044] (6) Inoculate the primary fermentation broth with Lactobacillus plantarum CGMCC NO.12849 and Enterococcus faecium CGMCC NO.9134 (1:1);

[0045] (7) inoculating the primary fermentation broth with Lactobacillus plantarum CGMCC NO.12849 and Lactobacillus reuteri CGMCC NO.19493 (1:1);

[0046] (8) Lactobacillus acidophilus CGMCC NO.5093 and Enterococcus faecium CGMCC NO.9134 (1:1) were inoculated into the primary fermentation liquid;

[0047] (9) inoculating the primary fermentation broth with Lactobacillus acidophilus CGMCC No.5093 and Lactobacillus reuteri CGMCC No.19493 (1:1);

[0048] (10) Enterococcus faecium CGMCC NO.9134 and Lactobacillus reuteri CGMCC NO.19493 (1:1) were inoculated into the primary fermentation broth;

[0049] (11) Inoculate the primary fermentation broth with Lactobacillus plantarum CGMCC NO.12849, Lactobacillus acidophilus CGMCC NO.5093, and Enterococcus faecium CGMCC NO.9134 (1:1:1);

[0050] (12) Inoculate the primary fermentation broth with Lactobacillus plantarum CGMCC NO.12849, Lactobacillus acidophilus CGMCC NO.5093, and Lactobacillus reuteri CGMCC NO.19493 (1:1:1);

[0051] (13) Lactobacillus acidophilus CGMCC NO.5093, Enterococcus faecium CGMCC NO.9134, and Lactobacillus reuteri CGMCC NO.19493 (1:1:1) were inoculated into the primary fermentation broth;

[0052] (14) The primary fermentation broth was inoculated with Lactobacillus plantarum CGMCC NO.12849, Lactobacillus acidophilus CGMCC NO.5093, Enterococcus faecium CGMCC NO.9134, and Lactobacillus reuteri CGMCC NO.19493 (1:1:1:1).

[0053] The antibacterial results of the above treatments are shown in Table 1. The results show that Scheme 13, i.e., inoculating the primary fermentation broth with Lactobacillus acidophilus CGMCC NO.5093, Enterococcus faecium CGMCC NO.9134 and Lactobacillus reuteri CGMCC NO.19493 (1:1:1), has the best antibacterial effect. However, considering the production cost, Scheme 6, i.e., inoculating the primary fermentation broth with Lactobacillus plantarum CGMCC NO.12849 and Enterococcus faecium CGMCC NO.9134 (1:1), is adopted.

[0054] Table 1 Comparison of antibacterial activity of different lactic acid bacteria alone and in combination

[0055]

[0056]

[0057] The fermentation liquid of Scheme 6 was centrifuged (8000 rpm, 10 min), the carrier was added to the supernatant, the inlet air temperature was 160°C, the outlet air temperature was 80°C, and the powder was spray-dried.

[0058] Lactobacillus plantarum CGMCC NO.12849 and Enterococcus faecium CGMCC NO.9134 were co-cultured in a 1:1 ratio and then spray-dried and powdered. After antibacterial test (spray-dried powder was diluted 100 times), it was found that the powder had good antibacterial effect on Gram-negative pathogenic bacteria Escherichia coli K88 (inhibition zone 22.03 mm) and Salmonella SL1344 (inhibition zone 20.83 mm).

[0059] Example 2 Screening of Gram-positive pathogenic bacteria

[0060] The strains involved in this embodiment are as follows:

[0061] Bacillus subtilis CGMCC NO.17348 is a strain of this experiment, named Bacillus subtilis 1-c-7, which was deposited in the General Microbiological Center of China Microbiological Culture Collection Administration on March 18, 2019, referred to as CGMCC, address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, with a deposit number of CGMCC NO.17348. See patent CN112574924A for details.

[0062] Bacillus subtilis CGMCC NO.25499 is a strain owned by our laboratory, named Bacillus subtilis BS-ZPW, classified and named: Bacillus subtilis, and deposited on August 8, 2022 at the General Microbiology Center of China Culture Collection Administration, referred to as CGMCC, address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, with a deposit number of CGMCC NO.25499. For details, see patent CN115181707A.

[0063] Bacillus subtilis CGMCC NO.13010 is a strain of our laboratory, named Bacillus subtilis 10#, which was deposited in the General Microbiological Center of China Microbiological Culture Collection Committee on September 19, 2016, address: No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, classified and named: Bacillus subtilis, and the deposit number is CGMCC NO.13010. For details, see patent CN118667706A.

[0064] The composite bacterial agent of Example 1 was compounded with other bacterial species to screen strains that can inhibit Gram-positive pathogenic bacteria for combination, and a yin-yang dual-antibacterial microecological powder was prepared. The existing bacterial powders that inhibit Gram-positive pathogenic bacteria in the laboratory were screened, and the antibacterial properties were compared and analyzed.

[0065] 1. Culture medium preparation:

[0066] (1) The seed culture medium is LB medium, and the composition and content are as follows: peptone 10 g / L, yeast extract powder 5 g / L, sodium chloride 10 g / L.

[0067] (2) The composition and content of the fermentation medium in the upper tank are as follows: peptone 8 g / L, soybean cake powder 40 g / L, glucose 5 g / L, corn starch 40 g / L, yeast extract powder 3 g / L, potassium phosphate trihydrate 2 g / L, magnesium sulfate 0.25 g / L, manganese sulfate 0.3 g / L, light calcium carbonate 3 g / L, sodium chloride 2 g / L, pH 7.0±0.2.

[0068] 2. High-density fermentation method

[0069] (1) Primary seed liquid culture

[0070] Take single colonies of Bacillus subtilis CGMCCNO.17348, Bacillus subtilis CGMCCNO.25499, and Bacillus subtilis CGMCCNO.13010 from the plates, inoculate them into LB liquid culture medium, and culture them at 32° C. for 16-18 hours to obtain a primary seed solution.

[0071] (2) Secondary seed liquid culture

[0072] The first-level seed solution cultured in step 1 was inoculated into LB liquid culture medium with an inoculation amount of 2% and cultured at 32° C. overnight to obtain the second-level seed solution.

[0073] (3) Liquid fermentation

[0074] After the fermentation medium is sterilized, cooling water is passed to keep the temperature at 32°C, and the secondary seed liquid is inoculated at a 10% inoculation amount into a 100L fermentation tank containing 50L of liquid fermentation medium, and the dissolved oxygen is controlled at 30%-40%. The fermentation culture is carried out for 12-18h. After the fermentation is completed, the number of viable bacteria in the fermentation liquid is 3.7×10 10 CFU / mL.

[0075] (4) Spray drying

[0076] The fermentation liquid obtained in step (3) is filtered through a 40-mesh filter, a carrier is added to the filtrate, the inlet air temperature is 160° C., the outlet air temperature is 80° C., and the powder is spray-dried.

[0077] Antibacterial evaluation method:

[0078] The samples were diluted 100 times with sterile water, and the inhibitory diameters against Escherichia coli K88, Salmonella SL1344, Staphylococcus aureus 25923 and Clostridium perfringens 13124 were determined by the double-layer agar Oxford cup method.

[0079] Trial Groups:

[0080] Test group 1 was the composite bacterial agent of Lactobacillus plantarum CGMCC NO.12849 and Enterococcus faecium CGMCC NO.9134;

[0081] Experimental group 2 was the above composite bacterial agent with Bacillus subtilis CGMCCNO.17348 powder added in a ratio of 1:1;

[0082] Experimental group 3 was the above composite bacterial agent plus Bacillus subtilis CGMCCNO.25499 powder in a ratio of 1:1;

[0083] Experimental group 4 was the above-mentioned composite bacterial agent plus Bacillus subtilis CGMCCNO.13010 powder in a ratio of 1:1.

[0084] The antibacterial results of microecological powders with different formulations are shown in Table 2. The results show that the antibacterial effect of test group 3 is the best. Figure 1 As shown, the compound bacterial agents of Lactobacillus plantarum CGMCC NO.12849 and Enterococcus faecium CGMCC NO.9134 are compounded with Bacillus subtilis CGMCC NO.25499.

[0085] Table 2 Comparison of antibacterial activity of microecological powders with different proportions

[0086]

[0087] Among them: 1 is a composite bacterial agent of Lactobacillus plantarum CGMCC NO.12849 and Enterococcus faecium CGMCC NO.9134; 2 is a microecological powder composed of a composite bacterial agent of Lactobacillus plantarum CGMCC NO.12849 and Enterococcus faecium CGMCC NO.9134 plus Bacillus subtilis CGMCC NO.17348 powder; 3 is a microecological powder composed of a composite bacterial agent of Lactobacillus plantarum CGMCC NO.12849 and Enterococcus faecium CGMCC NO.9134 plus Bacillus subtilis CGMCC NO.25499 powder; 4 is a microecological powder composed of a composite bacterial agent of Lactobacillus plantarum CGMCC NO.12849 and Enterococcus faecium CGMCC NO.9134 plus Bacillus subtilis CGMCC NO.13010 powder.

[0088] Example 3 Screening of the optimal ratio of antibacterial microecological powder

[0089] The invention discloses an antibacterial microecological powder, which is composed of the following raw materials: 1 part of a composite bacterial agent of Lactobacillus plantarum CGMCC NO.12849 and Enterococcus faecium CGMCC NO.9134, and 1 part of bacterial powder of Bacillus subtilis CGMCC NO.25499.

[0090] The invention discloses an antibacterial microecological powder, which is composed of the following raw materials: 7 parts of a composite bacterial agent of Lactobacillus plantarum CGMCC NO.12849 and Enterococcus faecium CGMCC NO.9134, and 3 parts of bacterial powder of Bacillus subtilis CGMCC NO.25499.

[0091] The invention discloses an antibacterial microecological powder, which is composed of the following raw materials: 10 parts of a composite bacterial agent of Lactobacillus plantarum CGMCC NO.12849 and Enterococcus faecium CGMCC NO.9134, and 1 part of bacterial powder of Bacillus subtilis CGMCC NO.25499.

[0092] The samples were diluted 100 times with sterile water, and the inhibitory diameters against Escherichia coli K88, Salmonella SL1344, Staphylococcus aureus 25923 and Clostridium perfringens 13124 were determined by the double-layer agar Oxford cup method.

[0093] The antibacterial results of the microecological powders with different proportions are shown in Table 3. The results show that the microecological powders prepared by the combination of the composite bacterial agent and the Bacillus subtilis agent in Example 1 in a ratio of 7:3 have the best antibacterial effect and are relatively low in cost. The minimum inhibitory concentration of various pathogens was determined under this formula, and the results are shown in Table 3. Figure 2 The microecological powder can be diluted to 8000 times (125μg / mL) for Escherichia coli K88, Salmonella SL1344 and Staphylococcus aureus 25923, and can be diluted to about 2000 times (500μg / mL) for Clostridium perfringens 13124.

[0094] Table 3 Comparison of antibacterial activity of microecological powders with different proportions

[0095]

[0096] Example 4 Study on the stability of antibacterial microecological powder

[0097] 1. High temperature resistance

[0098] The microecological powder was diluted 100 times with sterile water, one portion was used as a control, and three portions were placed in a water bath at 80°C, 90°C, and 100°C for 0.5 h; the last portion was placed in an autoclave and treated at 120°C for 0.5 h. After the sample was cooled, the antibacterial changes were detected. The results are shown in Table 4. Different temperature treatments did not significantly change the antibacterial activity of the microecological powder, indicating that the microecological powder is resistant to high temperatures.

[0099] Table 4 Comparison of antibacterial activity at different temperatures

[0100]

[0101]

[0102] 2. pH resistance

[0103] The microecological powder was diluted 100 times with sterile water, and adjusted to pH 2.0, 4.0, 6.0, 8.0, and 10.0 with 1 mol / L HCl and 1 mol / L NaOH, respectively. After standing at room temperature for 2 hours, the pH was adjusted to neutral, and the untreated microecological powder was used as a control to determine the antibacterial activity. The results are shown in Table 5, and it can be seen that the pH change has no obvious effect on the antibacterial activity of the microecological powder.

[0104] Table 5 Comparison of antibacterial activity at different pH treatments

[0105]

[0106] 3. Simulated gastrointestinal tolerance test

[0107] Simulated gastric fluid: Pepsin was dissolved in PBS (pH 3.0) to a final concentration of 1 g / L and sterilized by filtration using a 0.22 μm filter membrane.

[0108] Simulated intestinal fluid: Trypsin was dissolved in PBS (pH 8.0) to a final concentration of 1 g / L, and sterilized by filtration using a 0.22 μm filter membrane.

[0109] The microecological powder was digested by trypsin and pepsin respectively, and the antibacterial activity was determined in a water bath at 37° C. The results are shown in Table 6, which show that the microecological powder can tolerate the gastrointestinal tract.

[0110] Table 6 Simulated gastrointestinal tolerance results

[0111]

[0112]

[0113] 4.37℃ accelerated test

[0114] The microecological powder was placed in an aluminum foil bag, sealed and packaged, and placed in a 37°C incubator for 1 month, and the antibacterial activity was tested every 1 week. The results are shown in Table 7, and it can be seen that the antibacterial activity of the microecological powder at 37°C for 1 month did not change significantly, indicating that the microecological powder was relatively stable.

[0115] Table 7 Accelerated test results

[0116]

[0117] Example 5 Application of antibacterial microecological powder in pig farming

[0118] The experiment selected 72 healthy and disease-free piglets of the same breed, normal growth and development, and was divided into 6 groups on average, with 12 pigs in each group. The control group was fed with ordinary feed, and the experimental group was fed with different ratios of the antibacterial microecological powder in the ordinary feed. The commercially available group (the main components of the commercially available groups 1 and 2 were Bacillus subtilis) was fed with commercially available microecological preparations in the ordinary feed. The test period was 20 days. Feed 3 times a day and drink water 3 times a day. The production of the experimental pigs was observed at any time, and the disease was found to be treated in time.

[0119] Production performance measurement: record the feed intake of each group every day, observe and record the status of the test pigs, treat any diseases promptly, and keep records. Measurement indicators: measure daily weight gain, feed-to-weight ratio and diarrhea rate;

[0120] Feed-to-weight ratio = average daily feed intake / average daily weight gain;

[0121] Diarrhea rate = (number of piglets with diarrhea during the test period × number of test days) / (total number of test piglets × number of test days) × 100%.

[0122] The results are shown in Table 8 below:

[0123] Table 8 Production performance test results

[0124] Daily weight gain (g) Material weight ratio (%) Diarrhea rate (%) Control group 436 1.68 5.79 Experimental Group 1 509 1.41 2.74 Experimental Group 2 550 1.25 1.81 Experimental Group 3 524 1.33 2.33 Commercially available group 1 483 1.51 3.83 Commercially available group 2 471 1.57 4.01

[0125] As can be seen from Table 8 above, the daily weight gain of piglets in the experimental groups 1 to 3 and the commercial groups 1 to 2 were significantly higher than that in the control group, and the feed-to-weight ratio was lower. In addition, the diarrhea rates of piglets in the experimental groups 1 to 3 and the commercial groups 1 to 2 were significantly lower than that in the control group. However, the overall effect of the experimental groups was better than that of the commercial groups, among which the experimental group 2 had the best effect.

[0126] Example 6 Application of antibacterial microecological powder in chicken farming

[0127] The experiment selected 600 healthy three-yellow broiler chicks and randomly divided them into 6 groups. Each group had 5 replicates, 20 in each replicate. Among them, the control group was fed with ordinary feed, the experimental group was fed with different ratios of the antibacterial microecological powder in ordinary feed, and the commercially available group (the main components of commercially available groups 1 and 2 were Bacillus subtilis) was fed with commercially available microecological preparations in ordinary feed, and the trial period was 20 days. The feeding method was four-layer stepped free-range, with regular feeding twice a day, automatic drinking water, and the production of the experimental chickens was observed at any time, and the disease was found to be treated in time.

[0128] Production performance measurement: record the feed intake of each group every day, observe and record the status of the test chickens, treat any diseases promptly, and keep records. Measurement indicators: measure daily weight gain, feed-to-weight ratio and diarrhea rate;

[0129] Feed-to-weight ratio = average daily feed intake / average daily weight gain;

[0130] Diarrhea rate = (number of chickens with diarrhea during the test period × number of test days) / (total number of test chickens × number of test days) × 100%.

[0131] The results are shown in Table 9 below:

[0132] Table 9 Production performance test results

[0133]

[0134]

[0135] As can be seen from Table 9 above, the daily weight gain of broilers in the experimental groups 1 to 3 and the commercial groups 1 to 2 was significantly higher than that in the control group, and the feed-to-weight ratio was lower. In addition, the diarrhea rates of broilers in the experimental groups 1 to 3 and the commercial groups 1 to 2 were significantly lower than that in the control group. However, the overall effect of the experimental groups was better than that of the commercial groups, and the experimental group 1 had the best effect.

[0136] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein by equivalents. However, these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A microecological powder, characterized in that: The microecological powder comprises composite fermented bacterial powder and bacillus subtilis powder; the composite fermented bacterial powder is prepared from lactobacillus plantarum CGMCC NO.12849 and enterococcus faecium CGMCC NO.9134; and the bacillus subtilis powder is prepared from bacillus subtilis CGMCC NO.25499.

2. The microecological powder according to claim 1, characterized in that: In parts by mass, the composite fermentation bacteria powder and the Bacillus subtilis bacteria powder are prepared according to (1-10): (1-3).

3. The microecological powder according to claim 2, characterized in that: The composite fermentation bacteria powder and the Bacillus subtilis bacteria powder are prepared in a ratio of 7:3 by mass.

4. Use of the probiotic powder according to any one of claims 1 to 3 in the preparation of drugs or feeds for inhibiting Gram-negative bacteria and / or Gram-negative bacteria.

5. The use according to claim 4, characterized in that The Gram-negative bacteria are Escherichia coli and Salmonella, and the Gram-positive bacteria are Staphylococcus aureus and Clostridium perfringens.

6. A microecological preparation containing the microecological powder according to any one of claims 1 to 3.

7. A feed containing the probiotic powder according to any one of claims 1 to 3.

8. Use of the microecological powder according to any one of claims 1 to 3 in the preparation of medicines or feeds for improving the growth performance of livestock and poultry.

9. The method for preparing the microecological powder according to any one of claims 1 to 3, characterized in that: Prepare as follows: (1) Lactobacillus plantarum CGMCC No. 12849 and Enterococcus faecium CGMCC No. 9134 were inoculated into a fermentation medium at a ratio of 1:1 for fermentation. After the fermentation was completed, the supernatant of the fermentation liquid after centrifugation was spray-dried into a powder; (2) inoculating Bacillus subtilis CGMCCNO.25499 into a fermentation medium for fermentation, and after the fermentation is completed, filtering the fermentation liquid and spray-drying it into a powder; (3) The bacterial powders obtained in step (1) and step (2) are mixed evenly according to a certain proportion to obtain the product.

10. The method according to claim 9, characterized in that After fermentation is completed, the number of viable bacteria in the fermentation liquid is (1~9)×10 10 CFU / mL.

Images