CN114736834B - Bacillus pumilus TS1 and application thereof - Google Patents
Bacillus pumilus TS1 and application thereof Download PDFInfo
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- CN114736834B CN114736834B CN202210555094.3A CN202210555094A CN114736834B CN 114736834 B CN114736834 B CN 114736834B CN 202210555094 A CN202210555094 A CN 202210555094A CN 114736834 B CN114736834 B CN 114736834B
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/02—Separating microorganisms from their culture media
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
Abstract
The application discloses bacillus pumilus TS1 and application thereof. Bacillus pumilus TS1 is acid and alkali resistant and has antibacterial property. The application of the bacillus pumilus TS1 in preparing the feed additive and the application of the bacillus pumilus TS1 in preparing the feed additive for relieving the damage caused by stress are protected. In the experiment, chickens were fed with feed supplemented with Bacillus pumilus TS1, and the weight of the chickens was significantly heavier than those fed with normal feed under the same conditions.
Description
Technical Field
The invention relates to the technical field of biology, in particular to bacillus pumilus TS1 and application thereof.
Background
Stress or stress response refers to a nonspecific systemic reaction of the body when stimulated by various strong factors (stressors). Stressors are environmental events or situations that place an adaptation requirement on the body and can cause objective changes in response to, steady state imbalance, also known as irritants or stimuli. Oxidative stress refers to the fact that when the body is subjected to various harmful stimuli, excessive high-activity molecules such as active oxygen and active nitrogen are generated in the body, and the scavenging capacity of cells to oxides is far from meeting the antioxidant capacity which is required to resist the existing oxidation degree, and an imbalance of an oxidation system and an antioxidant system can cause damage to cells and important biological molecules, so that the whole organism is potentially influenced. In the field of poultry farming, after the poultry is stimulated by a stress source, immunity and resistance can be reduced, and finally, the feed intake of the poultry is reduced, the growth and development are slowed down or stopped, the immunity is reduced, and even death is caused, so that huge economic loss is caused to the breeding industry.
The problem of stimulation of various stress sources to poultry growth is increasingly prominent when the large-scale application of intensive cultivation in poultry cultivation is achieved while good economic benefits are obtained. Such as: a crowded farming environment, capturing and driving away poultry, resulting in their frightening groups; heat, cold, etc., and other factors may be the source of stress. Many studies have shown that poultry is stimulated by various stress sources, resulting in decreased activity of superoxide dismutase, glutathione peroxidase, etc., and increased activity of Reactive Oxygen Species (ROS), NO, malondialdehyde (MDA) in the viscera or serum of the poultry tissue, thereby causing oxidative stress damage.
Stress in the poultry raising process is unavoidable, so that the method for relieving the damage caused by the stress is significant. Since stress is a non-specific injury to the animal body, there is no targeted drug. By adding proper bacillus pumilus into the feed, the damage to tissues and organs caused by the oxidative stress of poultry can be reduced. Therefore, finding a bacillus pumilus suitable for being added into feed has great significance in reducing tissue and organ damage caused by poultry oxidative stress.
Disclosure of Invention
The application provides bacillus pumilus which is added into feed for poultry to relieve tissue and organ injury caused by oxidative stress of the poultry.
In a first aspect, the present application provides a Bacillus pumilus (Bacillus pumilus) TS1, where the Bacillus pumilus TS1 has been deposited at the China center for type culture Collection (address: university of Wuhan, china) for a period of 2022, and a accession number is CCTCC No. M2022538.
Bacillus species, a genus of bacteria, are capable of forming spores (endospores). They have strong resistance to external harmful factors and wide distribution, and exist in soil, water, air, animal intestinal tracts and the like. The bacillus bacteria are large (4-10 μm), gram positive, strictly aerobic or facultative anaerobic, capsular bacilli. An important property of this genus of bacteria is the ability to produce spores that are particularly resistant to adverse conditions.
Bacillus pumilus (Bacillus pumilus) is a bacterium of the genus Bacillus, which is generally 0.6-0.7μm.times.2.0-3.0 μm, gram positive. Bacillus pumilus is often used for controlling wheat root rot and strawberry gray mold.
Alternatively, the 16s rDNA sequence of the Bacillus pumilus TS1 is as follows: SEQ ID NO:1, the bacillus pumilus TS1 has acid resistance and bile salt resistance and has antibacterial effect.
The advantages of Bacillus pumilus are specifically expressed as follows: (1) easy preservation and good stress resistance: the conditions of separation, culture and preservation are simple, the technical requirements on industrial chemical production are low, and the stable effect and the high activity in intestines and stomach can be kept; the spore produced by the self can resist the external severe environment (such as heat, ultraviolet rays, ionizing radiation, low pH (2-3) and the like); (2) In the production and cultivation process of poultry, the growth and development of the poultry can be promoted, the growth performance of the poultry can be improved, the nutrition requirement is simple, and the metabolism can be rapidly promoted; (3) Can also relieve the damage of tissue viscera caused by poultry oxidative stress or pathogenic bacteria infection. Bacillus pumilus has the advantages of easy preservation, high active ingredients and the like, and is worthy of being popularized and applied in a large scale.
Optionally, the bacillus pumilus TS1 is isolated from yak manure.
In a second aspect, the application provides an application of bacillus pumilus TS1 in preparing a feed additive.
The poultry industry mainly regulates intestinal flora through feed additives, thereby improving intestinal health. Although some progress has been made, the actual production effect is not the same in consideration of the type of additive, the kind of poultry, the feeding environment, and the like. Bacillus pumilus (Bacillus pumilus) is a feed additive that has unique advantages over other feed additives in animal production and is therefore widely used.
Optionally, the feed additive is in the form of freeze-dried powder.
The freeze-dried powder is prepared by freezing water in the liquid medicine in advance by adopting a vacuum freeze-drying method of a freeze dryer, and sublimating the frozen water in the liquid medicine under a vacuum sterile environment to obtain freeze-dried powder.
Optionally, the content of bacillus pumilus TS1 in the freeze-dried powder is more than or equal to 10 9 CFU/g。
Alternatively, the lyophilized powder may be used in an amount up to 1 g/day.
In a third aspect, the application provides an application of bacillus pumilus TS1 in preparing a feed additive for relieving damage caused by stress.
The emergency response is that the poultry and livestock are stimulated by a stress source to cause allergic reaction of sensitive organisms. Such as crowded farming environments, capturing and driving away poultry, resulting in their frightening groups; heat, cold, etc., and other factors may be the source of stress.
Oxidative stress refers to the fact that when the body is subjected to various harmful stimuli, excessive high-activity molecules such as active oxygen and active nitrogen are generated in the body, and the scavenging capacity of cells to oxides is far from meeting the antioxidant capacity which is required to resist the existing oxidation degree, and an imbalance of an oxidation system and an antioxidant system can cause damage to cells and important biological molecules, so that the whole organism is potentially influenced. Many researches at present show that under the stimulation of various stress sources, the activities of superoxide dismutase, glutathione peroxidase and the like in the viscera or serum of poultry tissues are reduced, and the activities of Reactive Oxygen Species (ROS), NO and Malondialdehyde (MDA) are increased, so that oxidative stress damage is caused, and finally, the feed intake of the poultry is reduced, the growth and development are slowed down or stopped, the immunity is reduced, even death is caused, and huge economic losses are caused for the breeding industry.
Because the stress is nonspecific damage to animal organisms, no specific medicine exists, and the stress damage can be relieved by adding proper feed additives to improve the health of poultry organisms, enhance the immunity, improve the raising environment and the like. Wherein TS1 belongs to bacillus pumilus and belongs to microorganisms which are allowed to be added by feed additives.
Drawings
In order to more clearly illustrate the technical solutions of the present application, the drawings that are needed in the embodiments will be briefly described below, and it will be obvious to those skilled in the art that other drawings can be obtained from these drawings without inventive effort.
FIG. 1 shows the BLAST results of 16S rDNA sequences at NCBI;
FIG. 2 is an agarose electrophoresis diagram of the PCR product of Bacillus pumilus TS 1;
FIG. 3 shows the result of viability of Bacillus pumilus TS1 in LB medium at pH=7.12, 6.04, 5.03, 4.06 and 3.02 for 24 hours (wherein the abscissa indicates the pH of the LB medium, and the ordinate indicates the viability of Bacillus pumilus TS 1);
FIG. 4 shows the results of viability of Bacillus pumilus TS1 in LB medium with bile salt concentration of 0.05%, 0.1%, 0.2% and 0.3% for 24 hours (wherein the abscissa indicates the bile salt concentration of LB medium, and the ordinate indicates the viability of Bacillus pumilus TS 1);
FIG. 5 is a zone of inhibition of Bacillus pumilus TS1 against gram-negative bacteria (wherein the left panel is E.coli and the right panel is Salmonella);
FIG. 6 is a zone of inhibition of Bacillus pumilus TS1 against gram-positive bacteria (wherein the left panel is Streptococcus and the right panel is Staphylococcus aureus);
FIG. 7 shows the effect of Bacillus pumilus TS1 on chicken body weight (wherein the abscissa indicates days of feeding, and the ordinate indicates chicken body weight; the dark color in the bar graph indicates control group, the light color indicates experimental group; and P < 0.05).
Detailed Description
Reference will now be made in detail to the embodiments, examples of which are illustrated in the accompanying drawings. When the following description refers to the accompanying drawings, the same numbers in different drawings refer to the same or similar elements, unless otherwise indicated. The embodiments described in the examples below do not represent all embodiments consistent with the present application. Merely as examples of systems and methods consistent with some aspects of the present application as detailed in the claims.
Example 1: separation and purification of Bacillus pumilus TS1
Small healthy yak dung samples from Gansu Shandan army horse-yard are clamped by forceps into a glass test tube filled with 5mL distilled water, and a mouth of the tube is sealed by a sealing film and boiled in boiling water for 10 minutes; taking 100 mu L of boiled supernatant to an EP tube filled with 5mL of LB liquid medium, and placing the mixture into a shaking table at 37 ℃ for 16-18h at 160-180 rpm; 100 mu L of the shaken bacterial solution is added into 5mL of liquid culture medium to shake the bacteria again for 16h.
Dipping and streaking the shaken bacterial liquid in an LB (liquid) plate, inversely culturing for 12-18h in a 37 ℃ incubator, picking single bacterial colony, inoculating the plate, culturing for 12-18h in the 37 ℃ incubator, purifying, picking the single bacterial colony again to 5mL of LB liquid medium, shaking for 16-18h to obtain bacterial liquid, mixing with the LB frozen stock solution with the same volume, and freezing and preserving at-80 ℃.
Example 2: identification of Bacillus pumilus TS1
Sequencing by using a 16S universal primer to determine the sequence of the isolated probiotics, wherein the 16S sequencing result is shown as SEQ ID NO. 1. The measured 16S rDNA sequences were analyzed by BLAST software against a database. As shown in FIG. 1, the results of BLAST of the 16S rDNA sequence on NCBI show that the sequence has homology of 99% or more with Bacillus pumilus.
In the PCR amplification reaction, 25. Mu.L of 2xTaq Plus Master Mix,2. Mu.L of primer 27F and 2. Mu.L of primer 1492R, 1-5. Mu.L of fresh bacterial liquid and a total of 50. Mu.L of redistilled water were mixed in a PCR tube with the fresh bacterial liquid, 16S universal primer and PCR enzyme. The amplification conditions for PCR were: pre-denaturation at 95℃for 3min; denaturation at 95℃for 15s, annealing at 50.6℃for 15s, elongation at 72℃for 100s, and cycling for 30-35 times; finally, the mixture is extended for 5min at 72 ℃.
And adding the PCR product into 1% agarose nucleic acid gel to carry out electrophoresis in 1xTAE electrophoresis liquid, and obtaining a 16S result after gel irradiation. As a result, as shown in FIG. 2, the target band appeared around 1670bp, and the negative control had no band.
Example 3: acid resistance test
Preparing an acidic culture medium: to 500mL of LB liquid medium, 6.4mL of 36.0% -38.0% hydrochloric acid solution was added, and the ph=7.0, 6.0, 5.0, 4.0 and 3.0 were adjusted at room temperature.
Shaking: taking 50 mu L of activated bacterial liquid by 1% inoculation, respectively adding the bacterial liquid into 4950 mu L of LB acidic liquid culture medium with pH of about 7.0, 6.0, 5.0, 4.0 and 3.0, taking 50 mu L of activated bacterial liquid, adding the bacterial liquid into 4950 mu L of normal LB liquid culture medium as a control, and simultaneously placing the bacterial liquid into a shaking table at 37 ℃ and a rotating speed of 160-180rpm for shaking for 16-18 hours.
Measuring absorbance: the absorbance (OD 600) of the bacterial solutions cultured in the acidic medium and the normal medium at 600nm was measured at shaking for 0h and 24h, respectively.
Calculating survival rate: according to beer-lambert law a= Kbc (a: absorbance; K: molar absorbance coefficient; b: light-absorbing layer thickness; c: concentration), bacteria have a maximum absorbance at a wavelength of 600nm and the bacterial concentration c=a/kb is proportional to the absorbance.
Survival = X 1 /X 0 ×100％(X 1 : the viable count of 24 hours of culture in an acid culture medium; x is X 0 : viable count of 0h in acid medium culture); the number of viable bacteria is expressed as bacterial concentration and the final survival rate can be expressed as: survival = a 1 /A 0 ×100％(A 1 : culturing in an acidic culture medium for 24 hours to obtain a light absorption value; a is that 0 : absorbance for 0h in acid medium
In the experimental acid resistance test, the pH values of the prepared LB culture solutions were specifically 7.12, 6.04, 5.03, 4.06 and 3.02, and the survival rates were as shown in fig. 3, and bacillus pumilus TS1 had good acid resistance properties, wherein the survival rate of bacillus pumilus TS1 was highest among the five groups at ph=7.12.
Example 4: alkali resistance test
Preparing a bile salt culture medium: 1.5g of ox gall salt was added to 500mL of LB liquid medium to make the content of ox gall salt 0.3%, and then the ox gall salt was diluted to 0.2%, 0.1% and 0.05% in a gradient.
Shaking: taking 50 mu L of activated bacterial liquid by 1% inoculation, respectively adding the bacterial liquid into 4950 mu L of culture solutions with different bile salt concentrations, then taking 50 mu L of activated bacterial liquid, adding the bacterial liquid into 4950 mu L of normal LB liquid culture medium as a reference, and simultaneously placing the bacterial liquid into a shaking table at the temperature of 37 ℃ and the rotating speed of 160-180rpm for shaking for 16-18 hours.
Measuring absorbance: OD600 values of culture bacteria liquid in bile salt culture liquids with different concentrations are respectively measured in shaking bacteria for 0h and 24 h;
calculating survival rate: survival = B 1 /B 0 ×100％(B 1 : culturing in bile salt culture medium for 24 hr to obtain absorbance; b (B) 0 : absorbance value for 0h in bile salt medium
The results of the alkali resistance test are shown in FIG. 4, and the Bacillus pumilus TS1 has good acid resistance. The 24-hour survival rate was higher than 60% in LB medium with 0.05%, 0.1%, 0.2% and 0.3% bile salt concentration, and the survival rate was more than 90% in LB medium with 0.05% bile salt concentration, and as the concentration was increased, the survival rate was decreased.
Example 5: in vitro bacteriostasis test of Bacillus pumilus TS1 on gram-negative bacteria
Culturing pathogenic bacteria: culturing escherichia coli and salmonella, recovering and activating TS1, and shaking the escherichia coli and salmonella synchronously with TS1 to obtain activated bacterial liquid;
dilution of pathogenic bacteria: taking five 10mL centrifuge tubes, adding 4.5mL sterile PBS into each tube, adding 500 mu L of bacterial liquid into a first tube, mixing uniformly, then taking 500 mu L of diluted bacterial liquid in the first tube, adding into a second tube, mixing uniformly … …, and so on for gradient dilution to obtain the bacterial liquid with the concentration of 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 Is a pathogenic bacteria liquid.
Paving: after the LB nutrient agar plate is blown aside by an alcohol lamp of an ultra-clean bench for 5-10 minutes, diluted escherichia coli and salmonella bacteria liquid are poured into a plate, and after the diluted escherichia coli and salmonella bacteria liquid are mixed uniformly, the redundant escherichia coli and salmonella bacteria liquid is sucked out, a plate cover is opened in the ultra-clean bench and is placed aside by the alcohol lamp for blowing for about 20 minutes.
Punching: after the flat plate is dried, punching small holes with the diameter of 5-6mm and the edge of which is neat in the flat plate by using a puncher;
culturing: taking 50-80 mu L of TS1 activated bacterial liquid into the hole, and placing the bacterial liquid in a constant temperature oven at 37 ℃ for culturing for 18-20 h after the bacterial liquid is completely absorbed.
As shown in FIG. 5, the bacillus pumilus TS1 has a good antibacterial effect on gram-negative bacteria.
Example 6: in vitro bacteriostasis test of Bacillus pumilus TS1 on gram-positive bacteria
Culturing pathogenic bacteria: culturing streptococcus and staphylococcus aureus, recovering and activating TS1, and shaking with TS1 to obtain activated bacteria liquid;
dilution of pathogenic bacteria: taking five 10mL centrifuge tubes, adding 4.5mL sterile PBS into each tube, adding 500 mu L of bacterial liquid into a first tube, mixing uniformly, then taking 500 mu L of diluted bacterial liquid in the first tube, adding into a second tube, mixing uniformly … …, and so on for gradient dilution to obtain the bacterial liquid with the concentration of 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 Is a pathogenic bacteria liquid.
Paving: after the LB nutrient agar plate is blown aside by an alcohol lamp of an ultra-clean bench for 5-10 minutes, pouring diluted streptococcus and staphylococcus aureus bacterial liquid into a plate, sucking out redundant streptococcus and staphylococcus aureus bacterial liquid after uniform mixing, and opening a plate cover in the ultra-clean bench and placing the plate cover aside by the alcohol lamp for blowing for about 20 minutes.
Punching: after the flat plate is dried, punching small holes with the diameter of 5-6mm and the edge of which is neat in the flat plate by using a puncher;
culturing: taking 50-80 mu L of TS1 activated bacterial liquid into the hole, and placing the bacterial liquid in a constant temperature oven at 37 ℃ for culturing for 18-20 h after the bacterial liquid is completely absorbed.
As shown in FIG. 6, the bacillus pumilus TS1 has a good antibacterial effect on gram-positive bacteria.
Example 7: effect of feed feeding with TS1 on chicken body weight
20 chickens were randomly divided into two groups, a control group and an experimental group. TS1 is added into the feed of the experimental group, and the feed is continuously fed for 15 days, wherein the feed addition amount is 109 CFU/day; the control group was fed by the same method for 15 days without TS1 in the feed. Body weights of two groups of chickens were measured and recorded on days 6, 8, 10, 12, 14, 16, 18 and 20.
The weight of the chickens increased with the number of days, the group added with TS1 was significantly heavier than the control group without feed, and there was a significant difference between the two groups (×p < 0.05)
The foregoing detailed description of the embodiments is merely illustrative of the general principles of the present application and should not be taken in any way as limiting the scope of the invention. Any other embodiments developed in accordance with the present application without inventive effort are within the scope of the present application for those skilled in the art.
Sequence listing
<110> Ningxia Anrison Biotechnology Co.Ltd
Nanjing university of agriculture
<120> Bacillus pumilus TS1 and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1422
<212> DNA
<213> Bacillus pumilus (Bacillus pumilus)
<400> 1
ctcataaggt tacctcaccg acttcgggtg ttaaaactct cgtggtgtga cgggcggtgt 60
gtacaaggcc cgggaacgta ttcaccgcgg catgctgatc cgcgattact agcgattcca 120
gcttcacgca gtcgagttgc agactgcgat ccgaactgag aacagattta tgggattggc 180
taaaccttgc ggtctcgcag ccctttgttc tgtccattgt agcacgtgtg tagcccaggt 240
cataaggggc atgatgattt gacgtcatcc ccaccttcct ccggtttgtc accggcagtc 300
accttagagt gcccaactaa atgctggcaa ctaagatcaa gggttgcgct cgttgcggga 360
cttaacccaa catctcacga cacgagctga cgacaaccat gcaccacctg tcactctgtc 420
cccgaaggga aagccctatc tctagggttg tcagaggatg tcaagacctg gtaaggttct 480
tcgcgttgct tcgaattaaa ccacatgctc caccgcttgt gcgggccccc gtcaattcct 540
ttgagtttca gtcttgcgac cgtactcccc aggcggagtg cttaatgcgt tagctgcagc 600
actaaggggc ggaaaccccc taacacttag cactcatcgt ttacggcgtg gactaccagg 660
gtatctaatc ctgttcgctc cccacgcttt cgctcctcag cgtcagttac agaccagaga 720
gtcgccttcg ccactggtgt tcctccacat ctctacgcat ttcaccgcta cacgtggaat 780
tccactctcc tcttctgcac tcaagtttcc cagtttccaa tgaccctccc cggttgagcc 840
gggggctttc acatcagact taagaaaccg cctgcgagcc ctttacgccc aataattccg 900
gacaacgctt gccacctacg tattaccgcg gctgctggca cgtagttagc cgtggctttc 960
tggttaggta ccgtcaaggt gcgagcagtt actctcgcac ttgttcttcc ctaacaacag 1020
agctttacga tccgaaaacc ttcatcactc acgcggcgtt gctccgtcag actttcgtcc 1080
attgcggaag attccctact gctgcctccc gtaggagtct gggccgtgtc tcagtcccag 1140
tgtggccgat caccctctca ggtcggctac gcatcgtcgc cttggtgagc cattacccca 1200
ccaactagct aatgcgccgc gggtccatct gtaagtgaca gccgaaaccg tctttcatcc 1260
ttgaaccatg cggttcaagg aactatccgg tattagctcc ggtttcccgg agttatccca 1320
gtcttacagg caggttaccc acgtgttact cacccgtccg ccgctaacat ccgggagcaa 1380
gctcccttct gttcgctcga cttgcatgta ttaggcacgc cg 1422
Claims (7)
1. Bacillus pumilus TS1, wherein the Bacillus pumilus TS1 has a preservation number of CCTCC No. M2022538.
2. The bacillus pumilus of claim 1, wherein the 16s rDNA sequence of bacillus pumilus TS1 is: SEQ ID NO:1, the bacillus pumilus TS1 is acid-resistant and cholate-resistant and has antibacterial property.
3. The bacillus pumilus of claim 1, wherein bacillus pumilus TS1 is isolated from yak manure.
4. Use of the bacillus pumilus TS1 of claim 1 in the preparation of a feed additive.
5. The use according to claim 4, wherein the feed additive is in the form of a lyophilized powder.
6. The use according to claim 5, wherein the content of Bacillus pumilus TS1 in the freeze-dried powder is not less than 10 9 CFU/g。
7. The use according to claim 5, wherein the lyophilized powder is used in an amount of up to 1 g/day.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102586144A (en) * | 2012-02-09 | 2012-07-18 | 中国科学院南海海洋研究所 | Bacillus pumilus, probiotics preparation and preparation method and application thereof |
| CN102911901A (en) * | 2012-10-25 | 2013-02-06 | 青岛蔚蓝生物集团有限公司 | Bacillus pumilus strain and application thereof |
| JP2014096996A (en) * | 2012-11-13 | 2014-05-29 | Tokyo Univ Of Agriculture & Technology | Novel bacillus nitrogen fixation bacterium, plant growth accelerator, and plant cultivation method |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102586144A (en) * | 2012-02-09 | 2012-07-18 | 中国科学院南海海洋研究所 | Bacillus pumilus, probiotics preparation and preparation method and application thereof |
| CN102911901A (en) * | 2012-10-25 | 2013-02-06 | 青岛蔚蓝生物集团有限公司 | Bacillus pumilus strain and application thereof |
| JP2014096996A (en) * | 2012-11-13 | 2014-05-29 | Tokyo Univ Of Agriculture & Technology | Novel bacillus nitrogen fixation bacterium, plant growth accelerator, and plant cultivation method |
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