CN107723267B - A kind of piglet source bacillus amyloliquefaciens and application thereof - Google Patents

Abstract

The invention discloses a piglet source bacillus amyloliquefaciens and application thereof in animal and plant production. The Bacillus amyloliquefaciens provided by the invention is Bacillus amyloliquefaciens HCoB1. Experiments prove that the Bacillus amyloliquefaciens HCoB1 has an antibacterial effect, and can be used for adjusting the microecological balance of animals, preventing animal diseases, reducing the disease probability of the animals and improving the growth performance of the animals. The microbial preparation HCoB1 prepared by the invention has very important significance for reducing the use of antibiotics, improving the quality of agricultural and pastoral products and protecting the environment, is beneficial to promoting the sustainable development of agriculture, and has better application prospect.

Description

A kind of piglet source bacillus amyloliquefaciens and application thereof

technical field

The invention relates to a bacillus amyloliquefaciens derived from piglet intestines, which can inhibit various intestinal pathogenic bacteria and plant pathogenic fungi.

Background technique

Antibiotics can reduce the probability of animal disease, improve animal growth performance, and improve the economic benefits of the breeding industry. However, with the widespread and excessive use of antibiotics, there have been problems such as pathogenic bacteria resistance, animal gastrointestinal micro-ecosystem disorder, and antibiotic residues in livestock products. , Animal excrement polluting the environment and other issues. Therefore, in order to keep animal production benefits from animal husbandry from being greatly affected and at the same time reduce the use of antibiotics, it is necessary to find effective substances that can replace or reduce the use of antibiotics. More and more probiotics have been reported to regulate the microecological balance of animals in the gastrointestinal tract, so as to prevent diseases and improve animal production performance. At the same time, as people have higher and higher quality requirements for animal products, it has become a trend to use safe, environmentally friendly and efficient probiotic microecological preparations to replace antibiotics. Bacillus in probiotics has attracted the attention of researchers and animal husbandry producers because of its high stress resistance, non-toxic and harmless, significant effect, and low cost.

Bacillus amyloliquefaciens is a Gram-positive bacterium widely distributed in nature. They can produce spores, have strong stress resistance, grow fast at the same time, are non-toxic to humans and animals, and do not pollute the environment. Understanding the biologically active substances produced by Bacillus amyloliquefaciens and rationally applying them in animal and plant production is of great significance for reducing the use of antibiotics and pesticides, improving the quality of agricultural and animal husbandry products and protecting the environment.

Contents of the invention

One object of the present invention is to provide a strain of Bacillus amyloliquefaciens HCoB1.

The preservation number of Bacillus amyloliquefaciens HCoB1 provided by the present invention is CGMCC No. 14599.

The classification of Bacillus amyloliquefaciens HCoB1 of the present invention is named Bacillus amyloliquefaciens, and the strain has been preserved in the General Microorganism Center of China Committee for Culture Collection of Microorganisms (CGMCC for short, address: Beijing, China) on September 8, 2017 No. 3, No. 1 Yard, Beichen West Road, Chaoyang District, Institute of Microbiology, Chinese Academy of Sciences, Zip Code 100101), and the deposit number is CGMCC No.14599.

Another object of the present invention is to provide a new application of the above-mentioned Bacillus amyloliquefaciens or its preparation or its fermented liquid or its metabolic liquid or its bacterial suspension or its culture liquid.

The present invention provides the application of the above-mentioned Bacillus amyloliquefaciens or its preparation or its fermented liquid or its metabolic liquid or its bacterial suspension or its culture liquid in antibacterial and/or bacteriostatic.

The present invention also provides the application of the above-mentioned Bacillus amyloliquefaciens or its preparation or its fermented liquid or its metabolic liquid or its bacterial suspension or its culture liquid in the preparation of antibacterial and/or bacteriostatic products.

In the above-mentioned application, the bacteria are bacteria and/or fungi; the bacteria can specifically be Micrococcus luteus (Micrococcus luteus NCIB8166), Enterotoxigenic Escherichia coli (Enterotoxigenic Escherichia coli) Enterotoxigenic Escherichiacoli K88), Salmonella enterica Serovar Typhimurium (Salmonella enterica Serovar Typhimurium G5787), methicillin-resistant Staphylococcus aureu (methicillin-resistant Staphylococcus aureu 1-1), Citrobacter rodentium (Citrobacter rodentium DBS100) or Proteus mirabilis (ATCC number 29906);

Specifically, the fungus may be Fusarium graminearum (Fusarium graminearum NRRL 31084) or Fusarium oxysporum (Fusarium oxysporum SRRC 1370).

The present invention also provides the application of the above-mentioned Bacillus amyloliquefaciens or its preparation or its fermented liquid or its metabolic liquid or its bacterial suspension or its culture liquid in regulating the micro-ecological balance of animals.

The present invention also provides the application of the above-mentioned Bacillus amyloliquefaciens or its preparation or its fermented liquid or its metabolic liquid or its bacterial suspension or its culture liquid in the preparation of products for regulating the micro-ecological balance of animals.

The present invention also provides the application of the above-mentioned Bacillus amyloliquefaciens or its preparation or its fermented liquid or its metabolic liquid or its bacterial suspension or its culture liquid in preventing animal diseases or reducing the disease probability of animals.

The present invention also provides the application of the above-mentioned Bacillus amyloliquefaciens or its preparation or its fermented liquid or its metabolic liquid or its bacterial suspension or its culture liquid in the preparation of products for preventing animal diseases or reducing the probability of animal diseases.

The present invention also provides the application of the above-mentioned Bacillus amyloliquefaciens or its preparation or its fermented liquid or its metabolic liquid or its bacterial suspension or its culture liquid in improving the growth performance of animals.

The present invention also provides the application of the above-mentioned Bacillus amyloliquefaciens or its preparation or its fermented liquid or its metabolic liquid or its bacterial suspension or its culture liquid in the preparation of products for improving the growth performance of animals.

In the above application, the Bacillus amyloliquefaciens is the above-mentioned Bacillus amyloliquefaciens HCoB1.

A final object of the invention is to provide a product.

The active ingredient of the product provided by the invention is Bacillus amyloliquefaciens or its preparation or its fermented liquid or its metabolic liquid or its bacterial suspension or its culture liquid;

The product has any function in the following 1)-5):

1) antibacterial and/or bacteriostatic;

2) Regulating the micro-ecological balance of animals;

3) Prevention of animal diseases;

4) Reduce the chance of animal disease;

5) Improving the growth performance of animals.

In the above products, the bacteria are bacteria and/or fungi; the bacteria can specifically be Micrococcus luteus, Enterotoxigenic Escherichia coli, Salmonella enterica Serovar Typhimurium, methicillin-resistant golden yellow Staphylococcus methicillin-resistant Staphylococcus aureu, Citrobacter rodentium or Proteus mirabilis; the fungus may specifically be Fusarium graminearum or Fusarium oxysporum.

The Bacillus amyloliquefaciens is the above-mentioned Bacillus amyloliquefaciens HCoB1.

The invention provides a Bacillus amyloliquefaciens HCoB1, the preservation number of which is CGMCC No.14599. The Bacillus amyloliquefaciens HCoB1CGMCC No.14599 proved by experiments has antibacterial effect, and has good inhibitory activity on bacteria and fungi, and can be used to adjust the micro-ecological balance of animals, prevent animal diseases, reduce the probability of animal diseases, Animal growth performance. Not only is it of great significance to reduce the use of antibiotics, improve the quality of agricultural and animal husbandry products and protect the environment, but also help to promote the sustainable development of agriculture, and has a good application prospect.

Description of drawings

Figure 1 shows the analysis results of metabolites (iturin) in the supernatant of HCoB1 fermentation broth.

Figure 2 is the analysis results of metabolites (surfactin) in the supernatant of HCoB1 fermentation broth.

Fig. 3 is the analysis result of the metabolite (spinebin) in the supernatant of the HCoB1 fermentation broth.

Figure 4 is the result of the antibacterial experiment of HCoB1 fermentation broth supernatant against Micrococcus luteus 8166.

Fig. 5 is the result of the antibacterial experiment of HCoB1 fermentation liquid supernatant to Escherichia coli ETEC.

Fig. 6 is the result of the antibacterial experiment of the supernatant of the HCoB1 fermentation liquid against Salmonella typhimurium.

Fig. 7 is the result of the antibacterial experiment of HCoB1 fermentation broth supernatant against methicillin-resistant staphylococcus aureus (MRSA).

Figure 8 shows the results of the antibacterial experiment of the HCoB1 fermentation broth supernatant against Citrobacter rodentium CB10.

Figure 9 shows the results of the antibacterial experiment of the HCoB1 fermentation broth supernatant against Proteus mirabilis DRL2.

Figure 10 is the result of the antibacterial experiment of HCoB1 fermentation liquid supernatant to fungus. The left picture shows the results of the antibacterial experiment on Fusarium graminearum; the right picture shows the results of the antibacterial experiment on Fusarium oxysporum.

Preservation instructions

Bacteria name: Bacillus amyloliquefaciens

Latin name: Bacillus amyloliquefaciens

Strain number: HCoB1

Preservation institution: General Microbiology Center of China Committee for the Collection of Microorganisms

Depository institution abbreviation: CGMCC

Address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing

Deposit date: September 8, 2017

Registration number of the collection center: CGMCC No.14599

Detailed ways

The experimental methods used in the following examples are conventional methods unless otherwise specified.

The materials and reagents used in the following examples can be obtained from commercial sources unless otherwise specified.

The quantitative tests in the following examples were all set up to repeat the experiments three times, and the results were averaged.

Micrococcus luteus NCIB8166 (Micrococcus flavus NCIB8166) in the following examples is in the literature "Xiao H, Chen X, Chen M, Tang S, Zhao X, and Huan L.2004.Bovicin HJ50, anovellantibiotic produced by Streptococcus bovis HJ50. Microbiology 150:103-108.", publicly available from the Institute of Microbiology, Chinese Academy of Sciences.

ETEC K88 in the following examples is described in the literature "Zhang L, Zhang L, Zhan X, Zeng X, Zhou L, Cao G, Chen A, Yang C. Effects of dietary supplementation of probiotic, Clostridium butyricum, on growth performance, immune response, Intestinal barrier function, and digestive enzyme activity in broiler chickens challenged with Escherichiacoli K88.J Anim Sci Biotechnol.2016Jan 26; 7:3.doi:10.1186/s40104-016-0061-4", the public can download from Chinese Academy of Sciences Microbiology obtained by the Institute.

The methicillin-resistant Staphylococcus aureus 1-1 in the following examples is in the literature "Wang J, Zhang L, Teng K, Sun S, Sun Z, Zhong J. Cerecidins, novel lantibiotics from Bacillus cereus with potent antimicrobial activity .Appl.Environ.Microbiol.2014,80(8):2633", and the public can obtain it from the Institute of Microbiology, Chinese Academy of Sciences.

Embodiment 1, the isolation and identification of HCoB1 bacterial strain

1. Isolation of HCoB1 strain

HCoB1 was isolated from the colon of healthy piglets aged 1.5 months. The specific separation method is as follows: take the contents of the colon of one and a half months old healthy piglets after dissection, dilute it with normal saline according to 10 ^-1 to 10 ^-6 , and spread it on the LB culture plate (the formula of 1L medium is pancreas Peptone 10g, yeast extract 5g, sodium chloride 10g, 1.5% agar powder, distilled water to 1L, autoclaved). After the culture plate was cultured in a 37°C incubator for 1 day, pick out the single colony with irregular edges and put it in liquid LB medium (1L medium formula is tryptone 10g, yeast extract 5g, sodium chloride 10g, distilled water to volume to lL, autoclaved). After overnight culture, the antibacterial activity of the monoclonal culture product against Micrococcus luteus 8166 was detected. It was found that the supernatant of the fermentation broth of one strain had obvious antibacterial activity, and it was named HCoB1.

2. Identification of HCoB1 strains

HCoB1 bacteria were collected after culture, and genomic DNA was extracted, and the 16S rRNA sequence was amplified with primers (27F: 5'-AGAGTTTGATCCTGGCTCAG-3' and 1492R: 5'-GGTTACCTTGTTACGACTT-3) to obtain PCR products. And sequence the PCR products.

Sequencing results show that the sequence of the PCR product is shown in Sequence 1, which was identified as Bacillus amyloliquefaciens after comparison and analysis on the NCBI website (https://blast.ncbi.nlm.nih.gov/Blast.cgi) .

After the above identification results, the name of the HCoB1 strain was determined to be Bacillus amyloliquefaciens, and its classification was named Bacillus amyloliquefaciens. CGMCC, Address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, Zip Code 100101), and the deposit number is CGMCC No.14599.

Example 2, Fermentation of HCoB1 bacterial strain and analysis of metabolites in its fermentation supernatant

1. Fermentation of HCoB1 strain

The HCoB1 bacterial strain isolated in Example 1 was inoculated in LB medium and fermented and cultured in a shaker (200 rpm) at 37 degrees for 24 hours to obtain HCoB1 fermentation broth, which was centrifuged at 12000 rpm for 2 min and collected. supernatant to obtain the supernatant of the HCoB1 fermentation broth. The HCoB1 fermentation broth supernatant was named microbial preparation HCoB1.

2. Analysis of metabolites in the supernatant of HCoB1 fermentation broth

After the supernatant of the HCoB1 fermentation broth was treated with ziptip, the molecular weight of the metabolites in it was identified and analyzed by mass spectrometry.

The results show that: the HCoB1 fermentation broth supernatant contains the following three lipopeptide active products with antibacterial activity: iturin (molecular weight is 1403.5 Da, corresponding to iturin with 14 carbon chain lengths; molecular weight is 1057.5 Da, corresponding to iturin with 15 carbon chain lengths; Figure 1), surfactin (molecular weight is 1008.6Da, corresponding to surfactant with 13 carbon chain lengths; molecular weight is 1022.6, corresponding to 14 carbon chains length of surfactin; molecular weight is 1058.6Da, corresponding to surfactant containing 15 carbon chain lengths (sodium peak); Fig. The molecular weight is 1449.7, which corresponds to the pintonin with 15 carbon chain lengths; the molecular weight is 1463.7Da, which corresponds to the pintonin with 16 carbon chain lengths; the molecular weight is 1477.7, which corresponds to the pintonin with 17 carbon chain lengths ; The molecular weight is 1491.7Da, corresponding to pintonin with 18 carbon chain lengths; the molecular weight is 1505.7, corresponding to pintonin homologues with 18 carbon chain lengths, and the sixth position of alanine is replaced by valine; The molecular weight is 1513.7Da, which corresponds to the 16-carbon-chain-length homologue (sodium peak), whose sixth alanine is replaced by valine; Figure 3).

Embodiment 3, the application of microbial preparation HCoB1 in antibacterial

(1) Application of microbial preparation HCoB1 in inhibiting bacterial growth

1. The inhibitory effect of microbial preparation HCoB1 on Micrococcus luteus

Using Micrococcus luteus NCIB8166 as indicator bacteria, the antibacterial activity of HCoB1 fermentation broth supernatant against Micrococcus luteus NCIB8166 was detected by "dig well diffusion method". Concrete steps are as follows: use medium containing S1 (solvent is water, solute and mass fraction thereof are as follows: 0.8% tryptone, 0.5% yeast extract, 0.5% glucose, 0.2% disodium bicarbonate, 0.5% sodium chloride, 0.1 % Tween 20) plate culture Micrococcus luteus NCIB8166, and placed in the refrigerator for about 20 days, pick a ring of vine yellow microspheres NCIB8166 in normal saline, suspend and mix, take 0.5ml suspension and add 20ml of melted Mix evenly in the S1 solid medium, pour it into a petri dish with a diameter of 90 mm, and lay it flat to obtain an indicator plate of Micrococcus luteus NCIB8166. After the plate is solidified, use a puncher to punch a small hole with a diameter of 5mm on each indicator plate, add 25ul of the HCoB1 fermentation broth supernatant that has been cultured for 8 hours and 24 hours respectively into the small hole, and use the same volume of LB medium as the As a positive control, place the plate in a 30-degree incubator for overnight culture.

The results showed that the supernatant of the HCoB1 fermentation broth had obvious antibacterial activity against Micrococcus luteus 8166, and the antibacterial activity increased with the prolongation of fermentation time (Figure 4).

2. The inhibitory effect of microbial preparation HCoB1 on enterotoxigenic Escherichia coli

Enterotoxigenic Escherichia coli (ETEC) is an important pathogen that causes diarrhea in humans and young animals, especially newborn piglets, calves, lambs and weaned piglets. morbidity and mortality are high. It is estimated that 50% of piglet deaths worldwide are caused by ETEC each year. In order to study whether the microbial preparation HCoB1 has an inhibitory effect on Escherichia coli ETEC, the following experiments were carried out:

Add the supernatant of the HCoB1 fermentation broth to the LB medium containing enterotoxigenic Escherichia coli ETEC K88 according to the volume fraction of 10% and 20%, respectively, to obtain the ETEC K88 culture system; C was cultured. The OD600 value of the ETEC K88 culture system was detected at different time points during the culture process, and the growth curve was drawn with the OD600 value as the vertical axis and the growth time as the horizontal axis to detect its growth. At the same time, no HCoB1 fermentation supernatant was added as a control.

The results showed that the supernatant of the HCoB1 fermentation broth had a significant inhibitory effect on the growth of enterotoxigenic Escherichia coli ETEC, and the higher the concentration of the fermentation broth, the more obvious the inhibitory effect ( FIG. 5 ).

3. The inhibitory effect of microbial preparation HCoB1 on Salmonella typhimurium

Salmonella typhimurium and ETEC are two major pathogens affecting the swine industry. Salmonella typhimurium can cause various infectious diseases in poultry and mammals, and can also cause infections in humans, especially infants. It is one of the main pathogens causing acute gastroenteritis. In order to study whether the microbial preparation HCoB1 has an inhibitory effect on Salmonella typhimurium, the following experiments were carried out:

The supernatant of the HCoB1 fermentation broth was added into the LB medium containing Salmonella enterica Serovar Typhimurium G5787 (ATCC number 14028) according to the volume fraction of 10% and 20% respectively, to obtain the Salmonella typhimurium culture system; The cultures were carried out in an automatic microbial growth analyzer Bioscreen C. The OD600 value of the Salmonella typhimurium culture system was detected at different time points during the culture process, and the growth curve was drawn with the OD600 value as the vertical axis and the growth time as the horizontal axis to detect its growth. At the same time, no HCoB1 fermentation supernatant was added as a control.

The results showed that the supernatant of the HCoB1 fermentation broth had a significant inhibitory effect on the growth of Salmonella typhimurium ( FIG. 6 ).

4. The inhibitory effect of microbial agent HCoB1 on methicillin-resistant Staphylococcus aureus (MRSA)

The supernatant of the HCoB1 fermentation broth was added to the LB medium containing methicillin-resistant Staphylococcus aureus 1-1 according to the volume fraction of 10% and 20%, respectively, to obtain methicillin-resistant Staphylococcus aureus Cocci culture system, cultured in the automatic microbial growth analyzer Bioscreen C after inoculation. The OD600 value of the methicillin-resistant Staphylococcus aureus culture system was detected at different time points during the culture process, and the growth curve was drawn with the OD600 value as the vertical axis and the growth time as the horizontal axis to detect its growth. At the same time, no HCoB1 fermentation supernatant was added as a control.

The results showed that the supernatant of the HCoB1 fermentation broth had a significant inhibitory effect on the growth of methicillin-resistant Staphylococcus aureus (Figure 7).

5. The inhibitory effect of microbial preparation HCoB1 on Citrobacter

Add the supernatant of the HCoB1 fermentation broth into the LB medium containing Citrobacter rodentium DBS100 (ATCC No. 51459) according to the volume fraction of 10% and 20%, respectively, to obtain the Citrobacter culture system. Cultures were performed in a growth analyzer Bioscreen C. The OD600 value of the Citrobacter culture system was detected at different time points during the culture process, and the growth curve was drawn with the OD600 value as the vertical axis and the growth time as the horizontal axis to detect its growth. At the same time, no HCoB1 fermentation supernatant was added as a control.

The results showed that the supernatant of the HCoB1 fermentation broth had a significant inhibitory effect on the growth of Citrobacter (Fig. 8).

6. The inhibitory effect of microbial preparation HCoB1 on Proteus mirabilis

The supernatant of the HCoB1 fermentation broth was added into the LB medium containing Proteus mirabilis (ATCC No. 29906) according to the volume fraction of 10% and 20%, respectively, to obtain the Proteus mirabilis culture system; Analyzer Bioscreen C for incubation. The OD600 value of the Proteus mirabilis culture system was detected at different time points during the culture process, and the growth curve was drawn with the OD600 value as the vertical axis and the growth time as the horizontal axis to detect its growth. At the same time, no HCoB1 fermentation supernatant was added as a control.

The results showed that the supernatant of the HCoB1 fermentation broth had a significant inhibitory effect on the growth of Proteus mirabilis ( FIG. 9 ).

(2) Application of microbial preparation HCoB1 in inhibiting fungal growth

1. The inhibitory effect of microbial preparation HCoB1 on Fusarium graminearum

The antibacterial activity of HCoB1 fermentation broth supernatant against Fusarium graminearum was determined by well digging diffusion method. The specific steps are as follows: culture Fusarium graminearum NRRL 31084 on a plate containing PDA medium (1L PDA medium formula is as follows: 200 grams of potatoes, 20 grams of glucose, 15 to 20 grams of agar, and distilled water to 1L). , inoculate the bacteria in the center of the plate and grow for 2-4 days, then use a puncher to punch a small hole with a diameter of 5mm on the plate around the colony circle, add 25ul of the HCoB1 fermentation broth supernatant that has been cultivated for 24 hours into the small hole, and at the same time With Bacillus subtilis B.subtilis 168 and Lactococcus lactis L.lactis H1 capable of producing nisin as controls, put the plate into a 30-degree incubator and cultivate it for 1-2 days.

The results showed that the supernatant of the HCoB1 fermentation broth had a significant inhibitory effect on the growth of Fusarium graminearum (Figure 10).

2. The inhibitory effect of microbial preparation HCoB1 on Fusarium oxysporum

The antibacterial activity of HCoB1 fermentation broth supernatant against Fusarium oxysporum was determined by well digging diffusion method. The specific steps are as follows: Use a plate containing PDA medium to culture Fusarium oxysporum SRRC 1370, inoculate the bacteria in the center of the plate and grow for 2-4 days, and then use a puncher to punch small holes with a diameter of 5mm on the plate around the colony circle , add 25ul of the HCoB1 fermentation broth supernatant cultured for 24 hours into the small wells, and at the same time, use Bacillus subtilis B.subtilis 168 and Lactococcus lactis L.lactis H1 that can produce nisin as controls, and put the plate into 30 degrees for cultivation box for 1-2 days.

The results showed that the supernatant of the HCoB1 fermentation broth had a significant inhibitory effect on the growth of Fusarium oxysporum (Figure 10).

sequence listing

<110>Institute of Microbiology, Chinese Academy of Sciences

<120> A kind of piglet-derived bacillus amyloliquefaciens and application thereof

<160>1

<170>PatentIn version 3.5

<210>1

<211>1408

<212>DNA

<213>Artificial Sequence

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tgaaggtttt cggatcgtaa agctctgttg ttagggaaga acaagtgccg ttcaaatagg 420

gcggcacctt gacggtacct aaccagaaag ccacggctaa ctacgtgcca gcagccgcgg 480

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cttgagtgca gaagaggaga gtggaattcc acgtgtagcg gtgaaatgcg tagagatgtg 660

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tgttaggggg tttccgcccc ttagtgctgc agctaacgca ttaagcactc cgcctgggga 840

gtacggtcgc aagactgaaa ctcaaaggaa ttgacggggg cccgcacaag cggtggagca 900

tgtggtttaa ttgaagcaac gcgaagaacc ttaccaggtc ttgacatcct ctgacaatcc 960

tagagatagg acgtccccctt cggggggcaga gtgacaggtg gtgcatggtt gtcgtcagct 1020

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Claims (10)

1. A strain of Bacillus amyloliquefaciens HCoB1, the preservation number of which is CGMCC No. 14599. 2. Application of Bacillus amyloliquefaciens or its preparation or its fermentation liquid or its metabolic liquid or its bacterial suspension or its culture liquid in the preparation of antibacterial and/or bacteriostatic products; said Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) is the bacillus amyloliquefaciens (Bacillus amyloliquefaciens) HCoB1 described in claim 1. 3. The application according to claim 2, characterized in that: the bacteria are bacteria and/or fungi. 4. application according to claim 3, is characterized in that: described bacterium is Micrococcus luteus (Micrococcusluteus), enterotoxigenic Escherichia coli (Enterotoxigenic Escherichia coli), Salmonella typhimurium (Salmonella enterica Serovar Typhimurium), armor-resistant Methicillin-resistant Staphylococcus aureu, Citrobacter rodentium or Proteus mirabilis; the fungus is Fusarium graminearum or Fusarium oxysporum. 5. Application of Bacillus amyloliquefaciens or its preparation or its fermentation liquid or its metabolic liquid or its bacterial suspension or its culture liquid in the preparation of products for regulating the microecological balance of animals; said Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) is the bacillus amyloliquefaciens (Bacillus amyloliquefaciens) HCoB1 described in claim 1. 6. Application of Bacillus amyloliquefaciens or its preparation or its fermentation liquid or its metabolic liquid or its bacterial suspension or its culture liquid in the preparation of products for preventing animal diseases or reducing the probability of animal diseases; Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) is Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) HCoB1 described in claim 1. 7. Application of Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) or its preparation or its fermentation liquid or its metabolic liquid or its bacterial suspension or its culture liquid in the preparation of products that improve the growth performance of animals; Bacillus amyloliquefaciens) is bacillus amyloliquefaciens (Bacillus amyloliquefaciens) HCoB1 described in claim 1. 8. A product whose active ingredient is Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) or its preparation or its fermented liquid or its metabolic liquid or its bacterial suspension or its culture liquid; The product has any function in the following 1)-5): 1) antibacterial and/or bacteriostatic; 2) Regulating the micro-ecological balance of animals; 3) Prevention of animal diseases; 4) Reduce the chance of animal disease; 5) Improve animal growth performance; The Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) is the Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) HCoB1 described in claim 1. 9. The product according to claim 8, characterized in that the bacteria are bacteria and/or fungi. 10. The product according to claim 9, characterized in that: the bacteria are Micrococcus luteus, Enterotoxigenic Escherichia coli, Salmonella enterica Serovar Typhimurium, nail resistant Methicillin-resistant Staphylococcus aureu, Citrobacter rodentium or Proteus mirabilis; the fungus is Fusarium graminearum or Fusarium oxysporum.

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