CN117683697A - Bacillus bailii Y01 and application thereof in bacteriostasis and improvement of animal growth performance - Google Patents
Bacillus bailii Y01 and application thereof in bacteriostasis and improvement of animal growth performance Download PDFInfo
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- CN117683697A CN117683697A CN202410137264.5A CN202410137264A CN117683697A CN 117683697 A CN117683697 A CN 117683697A CN 202410137264 A CN202410137264 A CN 202410137264A CN 117683697 A CN117683697 A CN 117683697A
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- 238000005303 weighing Methods 0.000 description 1
- 235000019786 weight gain Nutrition 0.000 description 1
- 230000004584 weight gain Effects 0.000 description 1
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Abstract
The invention discloses bacillus belgium Y01 and application thereof in bacteriostasis and improvement of animal growth performance, and relates to the technical field of screening of the bacillus belgium in bacteriostasis and promotion of animal growth, wherein bacillus belgium (Bacillus velezensis) Y01 is preserved in the China general microbiological culture collection center (CGMCC) for 1 month 4 days in 2024, the preservation number is CGMCC No.1.60195, the preservation address is North Chen Silu No.1, no. 3 in the area of the morning of Beijing city, and the classification name is bacillus belgium Bacillus velezensis; the bacillus belicus can antagonize staphylococcus aureus and staphylococcus epidermidis, has remarkable promoting effect on the growth performance of broiler chickens, can be used as a bacteriostatic agent to be applied to feed, replaces antibiotics, and reduces the generation of drug resistance.
Description
Technical Field
The invention relates to the technical field of screening bacillus belicus for inhibiting bacteria and promoting animal growth, in particular to bacillus belicus Y01 and application thereof in inhibiting bacteria and improving animal growth performance.
Background
Due to the large amount and improper use of antibiotics, serious negative effects are caused to the poultry industry, such as antibiotic residues in poultry products, intestinal microorganisms change, bacterial strain resistance, occurrence of multiple drug resistance and the like, and threat is also caused to public health safety. With the implementation of the policy of 'forbidden' at the feed end, reduced 'at the breeding end and limited' at the feed end in 2020, the 'alternate' product is rapidly developed. Wherein the enzyme preparation, the antibacterial peptide and the Chinese herbal medicine extract are mainly represented. However, these "tibody" products have limitations in the specific use process, such as the enzyme preparation does not achieve the use effect of antibiotics; the antibacterial peptide has small molecular weight, low natural content, difficult separation and purification and high chemical synthesis cost; and some Chinese herbal medicine extracts are relatively short in resources, unstable in drug effect and quality. Thus, it becomes particularly important to find an effective "tie-down" product.
Probiotics are considered to be one of the best "tibody" products, capable of maintaining the balance of flora in the animal's gut, proliferating beneficial bacteria, reducing enteropathogenic microorganisms and being successfully used as growth promoters for livestock and fish, mainly comprising lactic acid bacteria, yeasts and Bacillus (Bacillus) and the like. The bacillus has good stress resistance, strong stability, high growth speed, secretion of various enzymes (such as cellulase, amylase, protease, lipase and the like), can effectively inhibit the growth of pathogenic bacteria, has long storage period and easy processing and production, and is an ideal animal probiotic preparation production bacterium.
Bacillus belicus (Bacillus velezensis) is a novel strain of bacillus, and is a late-stage abnormal body of bacillus amyloliquefaciens (Bacillus amyloliquefaciens) in 2008, and is a gram-positive aerobic bacterium, the thallus is in a rod shape, the size of the thallus is 0.5 x (1.5-3.5) mu m, and the endospores are commonly existing in nature, such as air, soil, river water, sea, plant root systems, plant surfaces, animal intestinal tracts and the like. Bacillus beleiensis was originally isolated from a estuary named as Beleis river (Velez) in southern Spanish city-Ma Jiala by Ruiz-Garcinia, et al, and was therefore named Bacillus beleiensis.
In recent years, research on bacillus belgium at home and abroad has been mainly focused on plant disease control, such as: promoting plant growth, inhibiting infection of pathogenic bacteria, inducing plant system resistance, identifying antibacterial substances and gene cluster, antagonizing action mechanism, etc. However, in the aspect of prevention and control of animal diseases, there have been relatively few studies on bacillus bailii.
Therefore, how to screen bacillus belicus strains which can inhibit bacteria and promote animal growth is a problem which needs to be solved by the technicians in this field.
Disclosure of Invention
In view of the above, the invention provides bacillus beijerinus Y01 and application thereof.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
bacillus belicus Y01, wherein the Bacillus belicus (Bacillus velezensis) Y01 is preserved in China general microbiological culture collection center (CGMCC) with the preservation number of CGMCC No.1.60195 at the 1 st month and 4 th day of 2024, and the preservation address is the 1 st day and the 3 st day of the Beijing Chaoyang area, and is classified and named as Bacillus belicus Bacillus velezensis.
Preferably, the 16S rDNA sequence of the bacillus beijerinus Y01 is shown as SEQ ID NO.1.
As the same inventive concept as the technical scheme, the invention also claims the application of bacillus belicus Y01 in pathogen inhibition.
In the above technical scheme, the pathogenic bacteria are staphylococcus aureus and staphylococcus epidermidis.
As the invention concept same as the technical scheme, the invention also claims the application of the bacillus beijerinckii Y01 in improving the growth performance of animals.
In the above application, the bacillus belicus Y01 increases the average daily gain and feed to meat ratio of the animal.
As the same inventive concept as the technical scheme, the invention also claims a bacteriostatic agent comprising bacterial suspension or fermentation broth of bacillus bailii Y01; the active ingredients in the fermentation broth comprise amylase and protease.
As the invention conception same as the technical scheme, the invention also claims a feed for improving the growth performance of animals, comprising bacterial suspension or fermentation liquor of bacillus beijerinus Y01; the active ingredients in the fermentation broth comprise amylase and protease.
As the same invention conception as the technical scheme, the invention also claims a preparation method of fermentation broth of bacillus beziacillus Y01, which comprises the following steps: inoculating Bacillus bailii Y01 into LB liquid medium, shake culturing at 37deg.C 180r/min for 20h, centrifuging at 4deg.C 5000g for 10min, collecting supernatant, and filtering with 0.22 μm filter membrane to obtain sterile fermentation broth of Bacillus bailii Y01.
Compared with the prior art, the bacillus belicus (Bacillus velezensis) Y01 provided by the invention is sensitive to 21 common antibiotics, does not generate drug resistance, has good antagonism to staphylococcus, has the capability of producing amylase and protease, is used for hydrolyzing starch and protein components in feed, promotes digestion and absorption, has no hemolysis phenomenon, and is safe to broilers. The bacillus bailii bacterial suspension promotes the average weight gain of the broiler chicken test powder to be 6.57% (p is less than 0.05), has a certain growth promoting effect on the growth of the broiler chicken, is a novel probiotic strain with excellent performance, and has good development and application prospects.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the embodiments or the description of the prior art will be briefly described below, and it is obvious that the drawings in the following description are only embodiments of the present invention, and that other drawings can be obtained according to the provided drawings without inventive effort for a person skilled in the art.
FIG. 1 shows colony morphology of Bacillus bailii Y01 of example 1 of the present invention cultured on LB solid medium at 37℃for 16-24 h;
FIG. 2 is a gram stain photograph of Bacillus belicus Y01 provided in example 1 of the present invention under a 1000-fold optical microscope;
FIG. 3 is a map of the whole genome of Bacillus bailii Y01 according to example 1 of the present invention;
FIG. 4 is a 16S rDNA system-based evolutionary tree of Bacillus bailii Y01 provided in example 1 of the present invention;
FIG. 5 is a graph showing the bacteriostatic activity of Bacillus bailii Y01 suspension against staphylococci according to example 3 of the present invention;
FIG. 6 is a graph showing the bacteriostatic activity of Bacillus bailii Y01 fermentation broth against Staphylococcus according to the invention as provided in example 3;
FIG. 7 shows the protease-producing ability of Bacillus bailii Y01 provided in example 4 of the present invention;
FIG. 8 shows the amylase-producing ability of Bacillus bailii Y01 provided in example 5 of the present invention;
FIG. 9 is a photograph showing that Bacillus bailii Y01 according to example 6 of the present invention has no hemolysis;
FIG. 10 is a graph showing survival of broiler chickens fed continuously for 7 days with Bacillus bailii Y01 at different concentrations as provided in example 7 of the present invention;
FIG. 11 is a schematic illustration of a 1X 10 embodiment of the invention provided in example 7 10 CFU/mL bacillus belgium Y01 was fed continuously for 7 days with paraffin sections of heart, liver, spleen, lung and kidneys of broiler chickens; a is a paraffin section of the heart of the broiler chicken of the control group, B is a paraffin section of the liver of the broiler chicken of the control group, C is a paraffin section of the spleen of the broiler chicken of the control group, D is a paraffin section of the lung of the broiler chicken of the control group, E is a paraffin section of the kidney of the broiler chicken of the control group, F is a paraffin section of the heart of the broiler chicken of the test group, G is a paraffin section of the liver of the broiler chicken of the test group, H is a paraffin section of the spleen of the broiler chicken of the test group, I is a paraffin section of the lung of the broiler chicken of the test group, and J is a paraffin section of the kidney of the broiler chicken of the test group.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1 isolated culture and identification of Strain Y01
1. Isolation culture of Strain Y01
Weighing 0.5g of cecum of chicken, placing in LB liquid medium (peptone 10g/L, yeast extract 5g/L, naCl 10g/L, pH=7.0, 121 ℃ high pressure sterilization 15 min), shake culturing at 37 ℃ 180rpm for 16-24h. The bacterial liquid is diluted to 10 by the sterile physiological saline -1 、10 -2 、10 -3 、10 -4 、10 -5 And 10 -6 Each concentration gradient was uniformly spread with 100. Mu.L of LB solid medium (peptone 10g/L, yeast extract 5g/L, naCl 10g/L, agar powder)20g/L, pH=7.0, and autoclaving at 121deg.C for 15 min), culturing at 37deg.C for 16-24 hr, and separating and purifying bacteria according to morphological characteristics such as colony size, color, transparency, and edge. And then carrying out primary screening and multiple secondary screening on antagonistic bacteria by taking staphylococcus aureus as indicator bacteria to finally obtain a bacterial strain with strong antibacterial effect, and the bacterial strain is named as Y01.
2. Identification of Strain Y01
The bacterial colony characteristics, physiological and biochemical characteristics and molecular biological detection results of the strain Y01 are combined, and the strain is identified as bacillus beijerinus (Bacillus velezensis). The specific technical scheme is as follows:
(1) Colony characteristics and cell morphology characteristics
After the strain Y01 is cultured on LB solid medium for 24 hours at 37 ℃, the colony is rough, slightly yellow, opaque, irregular in edge and has the diameter of about 2-4mm (shown in figure 1). Single colonies growing on LB solid medium are picked up to prepare slides, gram stain and microscopic examination. As shown in FIG. 2, the gram-stained photograph under the optical microscope shows that Y01 cells are rod-shaped, straight or nearly straight, and are gram-positive bacteria.
(2) Identification of physiological and biochemical characteristics
And (3) selecting single bacterial colonies to inoculate in a bacteria micro-detection tube for measuring the physiological and biochemical indexes. The classification and judgment were carried out by referring to "Berger's bacteriology evaluation Manual" and "common bacterial System evaluation Manual", and the results are shown in Table 1, and it was preliminarily judged that the strain Y01 belongs to the genus Bacillus.
TABLE 1 physiological and biochemical test results of strain Y01
Note that: + indicates positive, -indicates negative.
(3) Strain Y01 whole genome sequencing
After shaking culture of the strain Y01 in LB liquid medium at 37 ℃ 180r/min for 6 hours, 8000g of the strain is centrifuged for 5min to collect thalli, genomic DNA is extracted, and after the DNA quality detection is qualified, sequencing analysis is carried out by the Haipirson biosciences Co. Libraries of different inserts were constructed using whole genome shotgun (Whole Genome Shotgun, WGS) strategy, sequenced using second generation sequencing technology (Next-Generation Sequencing, NGS) based on Illumina NovaSeq sequencing platform, while using third generation single molecule sequencing technology, sequencing analysis was performed on these libraries separately based on Oxford Nanopore ONT sequencing platform. 16S rDNA is selected from the whole genome of Y01 for BLAST analysis, and a standard strain with higher homology is selected and used for constructing a phylogenetic tree by using a MEGA7.0 and a Neighbor-Joining method.
The genome size of the strain Y01 is 3929778bp, the whole genome map is shown in FIG. 3, the G+C content is 46.50%, the total gene is 3868, the coding region occupies 88.57% of the genome, and the strain Y01 comprises 9 ribosomal RNA (rRNA) operons and 86 transfer RNA (tRNA) (shown in Table 2).
TABLE 2 genomic information for Strain Y01
The genomic sequence was aligned to the nucleic acid sequence database (Nucleotide Sequence Database, NT) to give the species information alignment TOP5 of the genome as shown in table 3.
TABLE 3 NT pool species information comparison of Strain Y01 TOP5
The nucleotide sequence of the 16S rDNA of the strain Y01 is shown as SEQ ID NO.1, and according to the comparison result of genome species information and the analysis result of the phylogenetic tree and the physiological and biochemical characteristics of the combined 16S rDNA, the strain Y01 is identified as bacillus beleins (Bacillus velezensis), and the result is shown as figure 4; the bacillus belgium (Bacillus velezensis) Y01 is preserved in the China general microbiological culture Collection center (CGMCC) for 1 month and 4 days of 2024, the preservation number is CGMCC No.1.60195, the preservation address is North Chen Xili No.1, 3 of the Beijing area, and the classification is named bacillus belgium Bacillus velezensis.
Example 2 detection of Strain Y01 antibiotic susceptibility
Dipping concentration of 1X 10 with sterile cotton swab 8 The CFU/mL Y01 bacterial suspension is coated on MH solid culture medium (MH broth powder 21g/L, agar powder 20g/L, and autoclaved at 121 ℃ for 15 min), after being placed for 5min, 24 drug sensitive paper sheets are attached to the surface of a flat plate, after standing culture for 18h at 37 ℃, the result is observed and the diameter of a bacteriostasis ring is measured. Sensitivity determination is made according to the American clinical and laboratory standards institute antibacterial drug sensitivity experimental criteria. Experiments were repeated 3 times. The results show that bacillus belgium is sensitive to 21 antibiotics such as midecamycin, clarithromycin, clindamycin, penicillin, norfloxacin, ampicillin, tobramycin, erythromycin, compound neonomine, kanamycin, polymyxin B, ciprofloxacin, streptomycin, oxacillin, minocycline, ofloxacin, ceftriaxone, chloramphenicol, spectinomycin, cefquinome, gentamicin and the like, and is resistant to ceftazidime, tetracycline and aztreonam. The results are shown in Table 4.
TABLE 4 detection of antibiotic susceptibility of Strain Y01
Note that: s: sensitivity; r: drug resistance.
EXAMPLE 3 inhibition of pathogenic bacteria by Strain Y01
The main pathogenic bacteria such as escherichia coli, acinetobacter baumannii, staphylococcus epidermidis and staphylococcus aureus are used as indicator bacteria. Regulating the pathogenic bacteria to 1×10 with sterile physiological saline 8 CFU/mL, then dipping the bacterial liquid by using a sterile cotton swab, respectively coating the bacterial liquid on an LB solid culture medium, standing for 10min, and then punching. At the same time, Y01 was adjusted to 1X 10 using sterile physiological saline 6 CFU/mL, 100. Mu.L was added dropwise to the wells of the solid medium. After 10min, the cells were incubated at 37℃for 16-24h. And observing whether an obvious bacteriostasis ring appears around the Y01, and measuring the diameter of the bacteriostasis ring. The results show that Y01 has remarkable antibacterial effect on staphylococcus aureus and staphylococcus epidermidis, andhas no remarkable antibacterial effect on Escherichia coli and Acinetobacter baumannii, and has antibacterial diameters of 25+ -1.07 mm and 20+ -0.69 mm on Staphylococcus aureus and Staphylococcus epidermidis respectively (as shown in figure 5).
Shake culturing strain Y01 in LB liquid medium at 37deg.C 180r/min for 20 hr, centrifuging at 4deg.C 5000g for 10min, collecting supernatant, and filtering with 0.22 μm filter membrane to obtain sterile fermentation broth of Bacillus bailii Y01. Then, 100. Mu.L of the fermentation broth was added dropwise to the wells of the above solid medium. After 10min, the cells were incubated at 37℃for 16-24h. And observing whether an obvious bacteriostasis ring appears around the Y01, and measuring the diameter of the bacteriostasis ring. The results show that Y01 has remarkable antibacterial effect on staphylococcus aureus and staphylococcus epidermidis, but has no remarkable antibacterial effect on escherichia coli and acinetobacter baumannii, and the antibacterial diameters of the Y01 on staphylococcus aureus and staphylococcus epidermidis are 23+/-1.25 mm and 19+/-0.78 mm (as shown in figure 6), so that the strain Y01 has great application potential in preventing and treating staphylococcus.
Example 4 protease production ability test of Strain Y01
Shaking culture of strain Y01 in LB liquid medium at 37deg.C 180r/min for 20 hr, and adjusting to 1×10 with sterile physiological saline 6 CFU/mL, then 10 μl was dropped vertically onto a protein medium (agar 20g/L, ph=7.0, autoclaved at 121 ℃ for 15min, cooled to 60 ℃, 20mL of pasteurized 100g/L skim milk was taken, mixed well, poured onto a plate), after 10min, cultured at 37 ℃ for 24h with standing, whether or not a distinct proteolytic loop appeared around the Y01 strain was observed, and the diameter of the proteolytic loop was measured to evaluate the protease producing ability of the Y01 strain. The results showed that Y01 had a very good ability to solubilize proteins, with a lytic circle diameter (D) of 23.50.+ -. 0.58mm and a colony diameter (D) of 12.25.+ -. 0.96mm (see FIG. 7).
Example 5 detection of the Amylase-producing ability of Strain Y01
Shaking culture of strain Y01 in LB liquid medium at 37deg.C 180r/min for 20 hr, and adjusting to 1×10 with sterile physiological saline 6 CFU/mL, then 10. Mu.L was dropped vertically onto a starch medium (peptone 10g/L, soluble starch 20g/L, agar 20g/L, pH=7.0, 121℃high)Autoclaving for 15 min), after 10min, stationary culture was performed at 37℃for 24h, whether or not a distinct starch hydrolysis ring was present around the Y01 strain (the starch hydrolysis ring was observed after iodine solution was added dropwise to the plate), and the diameter of the hydrolysis ring was measured to evaluate the amylase-producing ability of the Y01 strain. The results show that Y01 has a very good capacity to dissolve starch, with a ring diameter (D) of 26.50.+ -. 3.00mm and a colony diameter (D) of 13.50.+ -. 1.91mm (see FIG. 8).
EXAMPLE 6 detection of hemolysis of Strain Y01
After shaking culture of the strain Y01 in LB liquid medium at 37 ℃ for 20 hours at 180r/min, dipping Y01 on blood agar medium by using an inoculating loop, standing and culturing for 24 hours at 37 ℃, observing whether a hemolytic loop appears around Y01, and using staphylococcus aureus as a positive control. As shown in FIG. 9, the staphylococcus aureus showed a clear hemolytic loop on the blood agar medium, while Y01 had no hemolytic loop, indicating that the strain Y01 had a certain safety.
EXAMPLE 7 safety evaluation of Strain Y01
The 7-day-old broilers were arbitrarily divided into 5 groups, namely, group A, group B, group C, group D and group E, each group of 10 broilers. Wherein, the group A is a blank control group, and 1mL of sterile physiological saline is fed by mixing materials every day; group B, group C, group D and group E are test groups, and 1mL of the feed is mixed and fed for 1×10 each day 10 CFU/mL、1×10 9 CFU/mL、1×10 8 CFU/mL and 1X 10 7 CFU/mL of Y01 was fed continuously for 7 days, and whether the broiler was ill or dead was observed. The results show that the broilers in the test group and the blank control group can drink water normally, have good mental state, do not have adverse reaction, and do not have any morbidity or mortality phenomenon (as shown in figure 10).
After 7 days of continuous feeding, 3 broiler chickens are selected from the test group B, the test group D and the blank control group A respectively, blood is taken from veins, after standing and coagulating, 2000g of blood is centrifuged for 10min at 4 ℃, and serum is taken to measure physiological and biochemical indexes such as glutamic pyruvic transaminase (ALT), glutamic oxaloacetic transaminase (AST) and alkaline phosphatase (ALP). And 1mL of blood sample is taken from vein, collected in EDTA anticoagulation tube, and blood index such as white blood cell count (WBC), red blood cell count (RBC) and Hemoglobin (HGB) is measured by blood cell analyzer. The results show that after 7 days of continuous feeding, physiological and biochemical indexes such as glutamic pyruvic transaminase (ALT), glutamic oxaloacetic transaminase (AST) and alkaline phosphatase (ALP) of broiler chickens have no obvious change in blood indexes such as white blood cell count (WBC), red blood cell count (RBC) and Hemoglobin (HGB), and have no obvious difference compared with a control group (shown in tables 5 and 6).
TABLE 5 physiological and biochemical indicators of broiler chickens fed with strain Y01 at different concentrations for 7 days
TABLE 6 blood index of broiler chickens fed with strain Y01 at different concentrations for 7 days
Meanwhile, immediately dissecting the broiler chicken, observing the color and morphology of the viscera, checking whether the viscera are diseased, picking up organs such as heart, liver, spleen, lung and kidney, soaking and fixing the organs for 24 hours by using 4% formaldehyde solution, embedding the organs in paraffin, preparing slices, carrying out hematoxylin-eosin staining, observing pathological changes under a 400-time optical microscope, and photographing. As shown in fig. 11, the heart, liver, spleen, lung and kidney tissue structures of the broiler chickens of the test group (F-J in fig. 11) were not significantly different from those of the control group (a-E in fig. 11): the myocardium of the broiler chicken of the test group is not infiltrated by inflammatory cells, and has no phenomena of edema, congestion and hemorrhage; liver cells do not have degeneration necrosis, but local liver cells are consistent with the control group, and slight particle degeneration appears; spleen lymphocytes have no necrosis and inflammatory cell infiltration; pulmonary arterioles and capillaries are free from congestion, the interior of the secondary bronchus is free from inflammatory exudates, and the interstitial mass is free from edema and is widened; tubular epithelial cells were not necrotic, interstitial was decongested, but local tubular epithelial cells were consistent with the control group, with mild particulate degeneration. These results indicate that strain Y01 is safe for broiler chickens.
Example 8 Effect of Strain Y01 on broiler growth Performance
Will 120 1 day old broilers (male and female halves) were arbitrarily divided into 3 groups, control group, test I group and test II group, 6 replicates each, 10 replicates each. Wherein, the control group: mineral water is used for feeding every day; test group I: 1' -10 is used every day 8 CFU/mL Y01 bacterial suspension mixed material is fed; test group II: 1' -10 is used every day 7 CFU/mL Y01 bacterial suspension mixed material is fed. The test was continued for 25 days with daily feed per chicken in grams: before 10 days of age, the daily feed gram number of each chicken is day-age +2; the daily feed gram number of each chicken is 1 and the daily age is 11-20 days old; the daily feed gram number of each chicken is equal to the daily age of the chick at 21-25 days of age. The weight of the chicks in the 25-day trial was weighed and recorded daily and the feed to meat ratio was calculated.
The calculation formula is as follows: average daily feed intake (g/g) = (total feed weight-total recycle weight)/days of feeding; average body weight at the end of the trial (g/only) =total body weight at the end of the trial/number of surviving chickens at the end of the trial; average body weight at test (g/g) =total body weight at test/number of surviving chickens at test; end-average weight gain (g /) = end-average body weight-initial average body weight; average daily gain (g/day) =test end average weight gain/days of feeding; feed to meat ratio = average daily feed intake/average daily gain.
The results are shown in Table 7, and the average weight gain of the end of the broiler test of group I is 6.57% (p < 0.05), which is significantly different from that of the control group; and the meat ratio of the test group I is significantly lower than that of the control group (p < 0.05).
TABLE 7 Effect of Strain Y01 on broiler growth Performance (1-25 days)
Note that the lower case letters of the same column indicate significant differences (p < 0.05), and that no letter designation indicates insignificant differences (p > 0.05).
In the present specification, each embodiment is described in a progressive manner, and each embodiment is mainly described in a different point from other embodiments, and identical and similar parts between the embodiments are all enough to refer to each other.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (9)
1. The bacillus belgium Y01 is characterized in that bacillus belgium (Bacillus velezensis) Y01 is preserved in China general microbiological culture collection center (CGMCC) with the preservation number of CGMCC No.1.60195 at the 1 st month and 4 th year of 2024, and is classified and named as bacillus belgium Bacillus velezensis at the preservation address of Beicheng Chaoyang area North Xielu No.1.
2. The bacillus beleiensis Y01 according to claim 1, wherein the 16S rDNA sequence of bacillus beleiensis Y01 is shown in SEQ ID No.1.
3. Use of bacillus belgium Y01 according to claim 1 for inhibiting pathogenic bacteria.
4. The use according to claim 3, wherein the pathogenic bacteria are staphylococcus aureus and staphylococcus epidermidis.
5. Use of bacillus belgium Y01 according to claim 1 for improving growth performance in animals.
6. The use according to claim 5, wherein bacillus beijerinckii Y01 increases the average daily gain and feed to meat ratio of an animal.
7. A bacteriostat, characterized by comprising a bacterial suspension or fermentation broth of bacillus bailii Y01; the active ingredients in the fermentation broth comprise amylase and protease.
8. A feed for improving animal growth performance, which is characterized by comprising bacterial suspension or fermentation liquor of bacillus bailii Y01; the active ingredients in the fermentation broth comprise amylase and protease.
9. A preparation method of a fermentation broth of bacillus bailii Y01 is characterized by comprising the following steps: inoculating Bacillus bailii Y01 into LB liquid medium, shake culturing at 37deg.C 180r/min for 20h, centrifuging at 4deg.C 5000g for 10min, collecting supernatant, and filtering with 0.22 μm filter membrane to obtain sterile fermentation broth of Bacillus bailii Y01.
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