CN111454867B - Phoxinus lagowskii waxy bacillus strain and application thereof - Google Patents

Phoxinus lagowskii waxy bacillus strain and application thereof Download PDF

Info

Publication number
CN111454867B
CN111454867B CN202010463563.XA CN202010463563A CN111454867B CN 111454867 B CN111454867 B CN 111454867B CN 202010463563 A CN202010463563 A CN 202010463563A CN 111454867 B CN111454867 B CN 111454867B
Authority
CN
China
Prior art keywords
strain
phoxinus
lagowskii
application
phoxinus lagowskii
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN202010463563.XA
Other languages
Chinese (zh)
Other versions
CN111454867A (en
Inventor
张东鸣
陈玉珂
吴振超
王秋举
蔺丽丽
郭志欣
刘洪健
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Jilin Agricultural University
Tonghua Normal University
Original Assignee
Jilin Agricultural University
Tonghua Normal University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Jilin Agricultural University, Tonghua Normal University filed Critical Jilin Agricultural University
Priority to CN202010463563.XA priority Critical patent/CN111454867B/en
Publication of CN111454867A publication Critical patent/CN111454867A/en
Application granted granted Critical
Publication of CN111454867B publication Critical patent/CN111454867B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/07—Bacillus
    • C12R2001/085—Bacillus cereus
    • 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
    • C12N1/205—Bacterial isolates
    • 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
    • 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
    • A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/02—Bacterial antigens
    • A61K39/07—Bacillus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/52—Bacterial cells; Fungal cells; Protozoal cells
    • A61K2039/522—Bacterial cells; Fungal cells; Protozoal cells avirulent or attenuated
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
    • A61K2039/552—Veterinary vaccine

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Immunology (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Health & Medical Sciences (AREA)
  • Polymers & Plastics (AREA)
  • Medicinal Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Wood Science & Technology (AREA)
  • Biochemistry (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Biotechnology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Animal Husbandry (AREA)
  • General Chemical & Material Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Food Science & Technology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Birds (AREA)
  • Marine Sciences & Fisheries (AREA)
  • Insects & Arthropods (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Molecular Biology (AREA)
  • Physiology (AREA)
  • Mycology (AREA)
  • Epidemiology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Fodder In General (AREA)

Abstract

The invention discloses a phoxinus lagowskii waxy bacillus strain and application thereof. The strain is named as LSG-1, the microorganism preservation number is CGMCC No.19595, the preservation date is 20 days 04 months 2020, and the preservation unit is China general microbiological culture Collection center. The invention discloses application of the probiotic bacteria additive in preparation of aquatic animal feed additives, which keeps micro-state balance between animal organisms and microorganisms in intestinal tracts, and participates in substance metabolism in the intestinal tracts, so that the effects of resisting bacteria, promoting growth and improving the feed utilization rate are achieved, the growth performance of fish bodies is improved, and reference is provided for the probiotic bacteria additive of the aquatic animal feed.

Description

Phoxinus lagowskii waxy bacillus strain and application thereof
Technical Field
The invention relates to a bacillus strain and application thereof, belongs to the technical field of aquatic products, and particularly relates to a phoxinus lagowskii waxy bacillus strain and application thereof.
Background
Phoxinus lagowski (Dybowski,1869)) belongs to cyprinid, arotenoideae, phoxinus wild small-sized economic fish, common individuals in natural water areas are 30g-40g, the maximum individuals in artificial culture can reach more than 500g, the fish is tender in meat quality, delicious in meat taste, less in intramuscular spines and extremely rich in nutrition, is high-protein and high-fat fish, contains abundant essential amino acids and delicious amino acids in human body, and is high-quality animal protein. The development prospect of artificial cultivation of phoxinus lagowskii is very wide.
Currently, in China, probiotics used as feed additives mainly belong to the genus Bacillus, such as Bacillus licheniformis, Bacillus coagulans, Bacillus cereus, Bacillus subtilis, etc. Bacillus cereus is widely distributed, is commonly found in soil, dust and sewage, and is also common in vegetable foods and many cooked foods. Bacillus cereus is an aerobic gram-positive bacillus which can grow well under anaerobic condition, has the advantages of promoting the growth of animals, improving immunity, preventing diseases and the like, and is widely applied to various fields of feed, agriculture, aquaculture and the like as probiotics. In the aspect of animal production, the bacillus cereus microecological preparation is applied to the breeding industry. The bacillus cereus enters the digestive tract of an animal in a spore state to grow and breed, keeps the micro-state balance state between the animal body and microorganisms in the intestinal tract, and participates in the metabolism of substances in the intestinal tract, thereby achieving the effects of resisting bacteria, promoting growth and improving the utilization rate of feed.
Among the prior documents, there are few documents relating to the application of Bacillus cereus to aquatic products, and Bacillus cereus belonging to fish sources is almost absent.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide a phoxinus lagowskii strain which can improve the growth performance of fish bodies and provides a reference for a probiotic additive of aquatic animal feed.
The invention also aims to provide the application of the phoxinus lagowskii waxy bacillus strain in preparing the aquatic animal feed additive.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention provides a phoxinus lagowskii waxy Bacillus (Bacillus cereus) strain, which is named as LSG-1, the preservation number of the microorganism is CGMCC No.19595, the preservation date is 20 months and 04 months in 2020, the preservation unit is the China general microbiological culture preservation management center, and the preservation address is as follows: xilu No.1 Hospital No.3, Beijing, Chaoyang, North.
The invention provides the application of the phoxinus lagowskii waxy bacillus strain in preparing aquatic animal feed additives.
Further, the optimum concentration of the phoxinus lagowskii waxy bacillus strain is 1 × 108cfu/mL。
The invention provides the application of the phoxinus lagowskii waxy bacillus strain in preparing the aquatic animal feed probiotic additive.
The invention provides an application of the phoxinus lagowskii waxy bacillus strain in preparing a cold water fish feed additive.
The invention provides the application of the phoxinus lagowskii waxy bacillus strain in preparing the probiotic additive of the cold water fish feed.
The invention provides application of the phoxinus lagowskii waxy bacillus strain in preparing cold water fish vaccine immunomodulators or immunopotentiators.
The invention provides an application of the phoxinus lagowskii waxy bacillus strain in preparing cold water fish live vector vaccines.
The invention provides the application of the phoxinus lagowskii waxy bacillus strain in preparing phoxinus lagowskii feed additives.
The invention provides the application of the phoxinus lagowskii waxy bacillus strain in preparing phoxinus lagowskii feed additives with the functions of resisting bacteria, promoting growth and high feed utilization rate.
The invention provides a phoxinus lagowskii waxy Bacillus (Bacillus cereus) strain, which has a nucleotide sequence shown in any one of 1) to 4):
1) a nucleotide sequence shown as SEQ ID No. 1; or
2) A nucleotide sequence capable of hybridizing with the complementary sequence of SEQ ID No. 1; or
3) A nucleotide sequence having at least 97% or more homology with the nucleotide sequence shown in SEQ ID No. 1; or
4) A nucleotide sequence variant in which deletion, substitution or insertion of one or more bases is performed on the basis of the nucleotide sequence shown in SEQ ID No.1 and which still has a similar function or activity.
Further, phoxinus lagowskii strain has a nucleotide sequence having at least 99% or more homology with the nucleotide sequence shown in SEQ ID No. 1.
The invention provides a primer for amplifying a strain sequence of phoxinus lagowskii waxy bacillus, wherein an upstream primer sequence of the primer is shown as SEQ ID No. 2; the sequence of the downstream primer is shown as SEQ ID No. 3.
Compared with the prior art, the invention has the following beneficial effects:
the phoxinus lagowskii waxy bacillus strain provided by the invention is beneficial to keeping the animal organism and the microorganisms in the intestinal tract in a micro-state balance state, and simultaneously participates in the metabolism of substances in the intestinal tract, thereby achieving the effects of resisting bacteria, promoting growth and improving the utilization rate of feed; the concentration is 1X 108The optimum concentration of cfu/mL of the bacillus cereus is that the weight gain rate is about 1.3 times of that of a control group, and the specific growth rate, the viscera body ratio, the intestinal length index and the intestinal body index are all the best; improve the growth performance of the fish body and provide reference for the probiotic additive of the aquatic animal feed.
Drawings
FIG. 1 shows the colony morphology of Bacillus cereus (Bacillus cereus) in the examples;
FIG. 2 shows a phylogenetic tree of nucleotide sequences of 16S rRNA of 10 species constructed by the adjacency method (NJ) in the example;
FIG. 3 shows the effect of Bacillus cereus on the resistance of phoxinus lagowskii in the test examples; all data represent survival; lower case letters between different groups indicate significance of difference (P < 0.05).
Detailed Description
The invention will now be further described with reference to the accompanying drawings and specific embodiments.
Example 1
Separation and identification of strains:
(1) isolation of the Strain
Taking out intestinal contents of phoxinus lagowskii under aseptic condition, adding aseptic normal saline for dilution, taking 100 μ L of diluent with 3 optimal dilutions, coating on LB culture medium, and culturing in 37 deg.C constant temperature incubator for 24 h. Selecting dominant floras with different morphological characteristics for pure culture, selecting a single colony when the colony morphologically grows consistently, and preserving the single colony by using glycerol with the volume fraction of 40% for later use.
(2) Identification of strains
Performing morphological identification on the strain: the morphology of the colonies after purification was observed as shown in FIG. 1 and gram stained microscopic examination was performed.
Performing physiological and biochemical identification on the strain: the method is described in handbook of identification of common bacteria systems. The biochemical characteristics of the separated strains were measured using a bacterial biochemical micro-reaction tube, and a strain of bacillus licheniformis was used as a reference strain for the synchronization experiment, as shown in table 1.
TABLE 1 Biochemical identification of the strains
Figure BDA0002511752330000031
Figure BDA0002511752330000041
Note: "+" indicates positive; "-" indicates negative.
Performing molecular biological identification on the strain:
first, a DNA template was prepared, and the LSG-1 strain was inoculated on an LB plate and cultured overnight at 37 ℃. And inoculating a single colony into an LB culture solution, and performing shake culture at 37 ℃ for 24 hours. Taking 1mL of bacterial suspension, preparing DNA by using a DNA extraction kit, and using the DNA as a template of PCR reaction. Then, PCR amplification, cloning and sequencing of 16S rRNA were performed.
16S rRNA was amplified by PCR using universal primers: 27F: AGAGTTTGATCMTGGCTCAG and 1492R: TACGGYTACCTTGTTACGACT.
PCR reaction (50. mu.L): 2xTransTaq
Figure BDA0002511752330000043
25 mu L of HiFi PCR Supermix I, 3 mu L of DNA template, 1 mu L of upstream primer and downstream primer respectively, and 20 mu L of double distilled water.
PCR procedure: pre-denaturation at 94 ℃ for 5 min; at 95 ℃ for 10s, at 55 ℃ for 20s, at 72 ℃ for 1.5min, for 35 cycles; extension at 72 ℃ for 10 min.
The PCR product was purified and then subjected to sequencing by Jilin province Kuumei Biotech Co., Ltd, and the result is shown in SEQ ID No. 1.
Finally, a sequence analysis and phylogenetic tree of 16S rRNA was constructed. The 16S rRNA sequence of the determined strain LSG-1 is subjected to Blast search and analysis in GenBank, a Clustal W method is selected to carry out homologous sequence analysis and comparison with the 16S rRNA sequences of different species of related genera, and after the similarity degree of the LSG-1 sequence and a known sequence is measured, Phylogenetic analysis (Phylogenetic analysis) is carried out. The phylogenetic tree was constructed by the orthotopic binding method (Neighborjoin), and this experiment constructed 13 phylogenetic trees of strains, which were replicated 1000 times by the Bootstrap method (boottrap), and the reliability of the analysis results was measured by the Consistency index (Consistency in-dex, CI), as shown in fig. 2.
Carrying out drug sensitivity test on the strain: the screened LSG-1 strain is inoculated in LB culture solution, shaking culture is carried out for 16-18 h at 38 ℃, 100 mu L of bacterial suspension is taken and evenly coated on an LB culture medium plate, and 13 drug sensitive paper sheets (as shown in the following table 2) are pasted on the surface of a plate culture medium. And (5) culturing for 24 hours in a constant-temperature incubator at 38 ℃, measuring the diameter of the inhibition zone, and judging the drug sensitivity of the strain. The results were judged according to the manufacturer's criteria and are shown in Table 2.
TABLE 2 drug sensitivity test results for LSG-1 Strain
Figure BDA0002511752330000042
Figure BDA0002511752330000051
Note: "-" indicates no zone of inhibition; "S" means highly sensitive; "I" indicates moderate sensitivity; "R" represents drug resistance.
(3) Identification results
The research separates a strain LSG-1 from the intestinal tract of phoxinus lagowskii. Through gram staining, morphological characteristics and physiological and biochemical characteristics identification, the strain LSG-1 is a gram-positive bacterium and can produce spores, and the physiological and biochemical characteristics are basically consistent with those of the bacillus cereus. The 16S rRNA sequence determination of the separated LSG-1 strain and the construction of a phylogenetic tree show that the LSG-1 strain has 100 percent of homology with the bacillus cereus and is gathered on the phylogenetic tree. The LSG-1 strain is identified as bacillus cereus by integrating the morphological characteristics, physicochemical characteristics and 16S rRNA gene analysis results of bacteria, is stored in an aquatic organism laboratory of Jilin agriculture university and is then stored in China general microbiological culture collection center.
Effect test example 1
Application effect in phoxinus lagowskii cultivation
(1) Design of experiments
Phoxinus lagowskii is cultured in an aquarium of 110cm multiplied by 45cm multiplied by 60 cm. Bacillus cereus was cultured in the presence of 0(A0) and 1X 108 (A1)、1.5×108(A2)、2×108(A3)、2.5×108(A4) The concentrations of the components are respectively added into basic feed to prepare experimental feed. Each group of three repeated aquariums is provided with 30 fishes in each repeated aquariums. 8 in the morning of each day: 00-9: 00, 5 late: 00-6: 00 regularly feeding the fish in an amount of about 3% of the weight of the fish, and recording the fish every dayThe water temperature is controlled to be about 20 ℃ under the conditions of ingestion, death and morbidity, water is properly changed according to the water body condition, and the water changing amount is one third of the water body each time. Formal experiments are carried out after one week of temporary culture, and the experimental time is 60 days.
(2) Growth indicator determination
After the test formally starts, the initial weight of phoxinus lagowskii in the control group and the test group is respectively measured, after feeding for 60 days, the feeding is stopped for 24 hours, after appropriate anesthesia, the final weight is measured, and anatomical sampling is carried out. The relative weight gain, feed efficiency, specific growth rate, visceral volume ratio, intestinal length index and intestinal volume index were calculated for each group of fish.
The results are shown in Table 3, the concentration being 1X 108The fish Bacillus cereus has optimal concentration, weight increasing rate about 1.3 times of that of control group, and specific growth rate, viscera ratio, intestinal length index and intestinal body index all preferably, and the concentration is 1.5 × 108And 2X 108The fish-derived Bacillus cereus of (4) tends to decrease with increasing concentration. Among the prior documents, there are few documents relating to the application of Bacillus cereus to aquatic products, and Bacillus cereus belonging to fish sources is almost absent.
TABLE 3 influence of Bacillus cereus on growth performance of Lagowskiella
Figure BDA0002511752330000061
Effect test example 2
Phoxinus lagowskii cultivation disease-resistant effect
(1) Design of experiments
Phoxinus lagowskii is cultured in an aquarium of 110cm multiplied by 45cm multiplied by 60 cm. Bacillus cereus was cultured in the presence of 0(A0) and 1X 108 (A1)、1.5×108(A2)、2×108(A3) The concentrations of the components are respectively added into basic feed to prepare experimental feed. Each group of three repeated aquariums is provided with 30 fishes in each repeated aquariums. 8 in the morning of each day: 00-9: 00, 5 late: 00-6: 00 feeding at regular time, wherein the feeding amount is about 3% of the weight of the fish, the feeding, death and morbidity of the fish are recorded every day, the water temperature is controlled to be about 20 ℃, and the water is fed according to the water bodyThe water is properly changed, and the water changing amount is one third of the water body each time. The formal experiment is carried out after the temporary culture for one week, and the test time is 60 days.
After the feeding test is finished, 10 fish with similar weight are selected from each repeated test fish, and the challenge test is started after temporary culture for 3 days. Respectively adopting a disposable syringe according to the proportion of 1.8 multiplied by 106cfu/g injected dose of Aeromonas hydrophila was intraperitoneally injected (injection concentration was determined by preliminary experiments before challenge test). The control group was injected with an equal volume of saline intraperitoneally. After injection was complete, the test fish were returned to the marked aquarium for rearing. And observing the swimming condition and the death condition of the fish every 2h, counting the death condition (death number and death time) of the fish, and calculating the survival rate after the toxicity attack. The challenge test lasted 15 days. After the challenge test is finished, blood of living fish is collected through tail veins by a disposable syringe, the blood is centrifuged for 10min at 4000r/min, serum is separated, and relevant immunity indexes are detected.
(1) Survival rate after toxic material attack
The test results are shown in fig. 3, which shows that the disease resistance of phoxinus lagowskii can be remarkably improved by adding bacillus cereus into the feed. 1.5X 10 of feed additive8The disease resistance of cfu/g bacillus cereus phoxinus lagowskii is strongest.
(2) Serum immunity index after toxic material attack
The addition of the bacillus cereus in the feed can obviously (P is less than 0.05) improve the contents of complement C3, complement C4, immunoglobulin M and lysozyme in the serum of phoxinus lagowskii after challenge and the activity of acid phosphatase. Wherein the addition amount of Bacillus cereus is 1.5 × 108The immunity of cfu/g phoxinus lagowskii is highest.
TABLE 4 influence of Bacillus cereus on the serum complement C3, complement C4, IgM, acid phosphatase and lysozyme of phoxinus lagowskii
Figure BDA0002511752330000071
Note: all data are expressed as "mean ± sem" (n ═ 3). Different letters in the same column indicate significance of difference (P < 0.05).
The present invention is not limited to the above-described embodiments, and various changes and modifications of the present invention are intended to be included within the scope of the claims and the equivalent technology of the present invention if they do not depart from the spirit and scope of the present invention.
Sequence listing
<110> Jilin university of agriculture
General and chemical colleges of education and education
<120> phoxinus lagowskii waxy bacillus strain and application thereof
<130> BJ2000419CN
<141> 2020-05-27
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 925
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 1
cgctataatg caagtcgagc gaatggatta agagcttgct cttatgaagt tagcggcgga 60
cgggtgagta acacgtgggt aacctgccca taagactggg ataactccgg gaaaccgggg 120
ctaataccgg ataacatttt gaaccgcatg gttcgaaatt gaaaggcggc ttcggctgtc 180
acttatggat ggacccgcgt cgcattagct agttggtgag gtaacggctc accaaggcaa 240
cgatgcgtag ccgacctgag agggtgatcg gccacactgg gactgagaca cggcccagac 300
tcctacggga ggcagcagta gggaatcttc cgcaatggac gaaagtctga cggagcaacg 360
ccgcgtgagt gatgaaggct ttcgggtcgt aaaactctgt tgttagggaa gaacaagtgc 420
tagttgaata agctggcacc ttgacggtac ctaaccagaa agccacggct aactacgtgc 480
cagcagccgc ggtaatacgt aggtggcaag cgttatccgg aattattggg cgtaaagcgc 540
gcgcaggtgg tttcttaagt ctgatgtgaa agcccacggc tcaaccgtgg agggtcattg 600
gaaactggga gacttgagtg cagaagagga aagtggaatt ccatgtgtag cggtgaaatg 660
cgtagagata tggaggaaca ccagtggcga aggcgacttt ctggtctgta actgacactg 720
aggcgcgaaa gcgtggggag caaacaggat tagataccct ggtagtccac gccgtaaacg 780
atgagtgcta agtgttagag ggtttccgcc ctttagtgct gaagttaacg cattaagcac 840
tccgcctggg gagtacggcc gcaaggctga aactcaaagg aattgacggg ggcccgcaca 900
agcggtggag catgtggttt aattc 925
<210> 2
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 2
agagtttgat cmtggctcag 20
<210> 3
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 3
tacggytacc ttgttacgac t 21

Claims (2)

1. A phoxinus lagowskii waxy Bacillus strain (Bacillus cereus) is named as LSG-1, the microorganism preservation number is CGMCC No.19595, the preservation date is 20 days 04 months 2020, and the preservation unit is the China general microbiological culture preservation management center.
2. The use of a phoxinus lagowskii strain according to claim 1 in the preparation of a phoxinus lagowskii feed additive.
CN202010463563.XA 2020-05-27 2020-05-27 Phoxinus lagowskii waxy bacillus strain and application thereof Active CN111454867B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202010463563.XA CN111454867B (en) 2020-05-27 2020-05-27 Phoxinus lagowskii waxy bacillus strain and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202010463563.XA CN111454867B (en) 2020-05-27 2020-05-27 Phoxinus lagowskii waxy bacillus strain and application thereof

Publications (2)

Publication Number Publication Date
CN111454867A CN111454867A (en) 2020-07-28
CN111454867B true CN111454867B (en) 2022-04-01

Family

ID=71678005

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202010463563.XA Active CN111454867B (en) 2020-05-27 2020-05-27 Phoxinus lagowskii waxy bacillus strain and application thereof

Country Status (1)

Country Link
CN (1) CN111454867B (en)

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103966135B (en) * 2014-05-14 2016-03-09 北京鑫洋水产高新技术有限公司 A kind of bacillus cereus and uses thereof and feed and uses thereof
CN105754886B (en) * 2014-12-16 2019-07-12 朱奎 One plant of nontoxic bacillus cereus strain pBC-1 and its application
CN105132310B (en) * 2015-08-07 2019-09-03 哈尔滨动物生物制品国家工程研究中心有限公司 One plant of cold water fish probiotics Bacillus strain and application thereof
CN105039221B (en) * 2015-08-07 2019-04-16 中国农业科学院哈尔滨兽医研究所 One plant of cold water fish probiotics Arthrobacter strain and application thereof
CN105861389B (en) * 2016-05-30 2019-06-28 天津师范大学 For improving Bacillus strain and its screening technique and the application of aquatic livestock growth performance
CN106720942A (en) * 2016-12-15 2017-05-31 上海创博生态工程有限公司 A kind of additive for microbe feedstuff for improving Eriocheir sinensis meat and preparation method thereof
CN108048340B (en) * 2017-10-10 2021-05-04 盐城裕达饲料有限公司 Bacillus cereus and application thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
洛氏鱥肠道产酶益生菌的筛选及其粘附特性分析;吴振超;《中国优秀硕士学位论文全文数据库_农业科技辑》;20210315(第3期);第D052-185页 *

Also Published As

Publication number Publication date
CN111454867A (en) 2020-07-28

Similar Documents

Publication Publication Date Title
CN108251335B (en) Enterococcus faecalis HKF7 with lactic acid activity and screening culture method and application thereof
CN114908013B (en) Shewanella manshurica for producing DDP-IV inhibitor and application thereof
CN111733117B (en) Bacillus marinus for producing antibacterial peptide and fermentation method and application thereof
CN108522884B (en) Application of composite microecological preparation in preventing fat deposition of broiler chickens
CN105524855B (en) A kind of bacillus coagulans and its application with aquatic pathogenic bacterium antagonistic properties
CN111593010B (en) Lactobacillus plantarum and method for producing fermented feed by using same
CN113430141A (en) Probiotic for aquatic feed and preparation method of microbial inoculum thereof
CN111004741B (en) Deep-sea-derived campylobacter R29-2, microecological preparation and application thereof
CN114134083A (en) Bacillus belgii and application thereof
CN113528375B (en) Min swine bacillus subtilis capable of inhibiting growth of various pathogenic bacteria microorganisms
CN112538442B (en) Bacillus licheniformis and application thereof
CN104263683B (en) Bacillus pumilus 315 and its application with prebiotic effect
JP6785324B2 (en) Bacillus licheniformis NY1505 strain that mass-produces α-glucosidase inhibitors
CN113502243A (en) Lactobacillus plantarum GBW-LP001 capable of highly producing lactic acid and antibacterial agent alternative thereof and application
CN111454867B (en) Phoxinus lagowskii waxy bacillus strain and application thereof
CN112442461B (en) Halomonas meibomiae and application thereof in prawn culture water regulation
CN111925972B (en) Lactobacillus hilgardii and application thereof
CN116083272A (en) Clostridium butyricum and application of biological feed additive thereof
Ebnetorab et al. Isolation, biochemical and molecular detection of Bacillus subtilis and Bacillus licheniformis from the digestive system of rainbow trout (Oncorhynchus mykiss) and its inhibitory effect on Aeromonas hydrophila
CN111411053B (en) Bacillus subtilis JCL16 capable of synergistically producing three antibacterial metabolites and screening and application thereof
CN114891678A (en) Bacillus polymyxa CPL258 and screening and application thereof
CN112375711B (en) Alkaliphilic monad separated from prawn
CN112322544B (en) Lujie&#39;s bacillus for prawn culture
CN117070418B (en) Acetylmicrobacterium strain with high salt tolerance, microbial agent and application thereof
CN114672437B (en) Bacillus subtilis G-1 for producing organic acid and application thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant