CN108060102A - A kind of cibarium Wei Si Salmonellas and its application, screening calibration method - Google Patents
A kind of cibarium Wei Si Salmonellas and its application, screening calibration method Download PDFInfo
- Publication number
- CN108060102A CN108060102A CN201810093014.0A CN201810093014A CN108060102A CN 108060102 A CN108060102 A CN 108060102A CN 201810093014 A CN201810093014 A CN 201810093014A CN 108060102 A CN108060102 A CN 108060102A
- Authority
- CN
- China
- Prior art keywords
- salmonellas
- screening
- cibarium wei
- lactic acid
- acid bacteria
- 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.)
- Granted
Links
- 241000607142 Salmonella Species 0.000 title claims abstract description 66
- 238000012216 screening Methods 0.000 title claims abstract description 39
- 238000000034 method Methods 0.000 title claims abstract description 24
- 239000004461 grass silage Substances 0.000 claims abstract description 24
- 238000004519 manufacturing process Methods 0.000 claims abstract description 20
- 239000000654 additive Substances 0.000 claims abstract description 17
- 230000000996 additive effect Effects 0.000 claims abstract description 14
- 239000002253 acid Substances 0.000 claims abstract description 13
- 241000975185 Weissella cibaria Species 0.000 claims abstract description 7
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 72
- 241000894006 Bacteria Species 0.000 claims description 69
- 239000004310 lactic acid Substances 0.000 claims description 36
- 235000014655 lactic acid Nutrition 0.000 claims description 36
- 241000209046 Pennisetum Species 0.000 claims description 19
- 239000000243 solution Substances 0.000 claims description 19
- 244000025254 Cannabis sativa Species 0.000 claims description 16
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 14
- 238000012408 PCR amplification Methods 0.000 claims description 13
- 230000001580 bacterial effect Effects 0.000 claims description 13
- 238000000855 fermentation Methods 0.000 claims description 13
- 230000004151 fermentation Effects 0.000 claims description 13
- 241000931313 Hemarthria Species 0.000 claims description 11
- 238000000605 extraction Methods 0.000 claims description 11
- 235000015097 nutrients Nutrition 0.000 claims description 11
- 238000012360 testing method Methods 0.000 claims description 11
- 239000004459 forage Substances 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 10
- 239000007787 solid Substances 0.000 claims description 10
- 108020004465 16S ribosomal RNA Proteins 0.000 claims description 8
- 238000004458 analytical method Methods 0.000 claims description 8
- 239000012895 dilution Substances 0.000 claims description 7
- 238000010790 dilution Methods 0.000 claims description 7
- 238000007400 DNA extraction Methods 0.000 claims description 6
- 239000012530 fluid Substances 0.000 claims description 6
- 230000015784 hyperosmotic salinity response Effects 0.000 claims description 6
- 238000005259 measurement Methods 0.000 claims description 6
- 102000016938 Catalase Human genes 0.000 claims description 5
- 108010053835 Catalase Proteins 0.000 claims description 5
- 238000011156 evaluation Methods 0.000 claims description 5
- 238000010186 staining Methods 0.000 claims description 5
- 238000003860 storage Methods 0.000 claims description 5
- 238000006243 chemical reaction Methods 0.000 claims description 4
- 239000002054 inoculum Substances 0.000 claims description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- 238000000137 annealing Methods 0.000 claims description 3
- 230000000692 anti-sense effect Effects 0.000 claims description 3
- 238000010876 biochemical test Methods 0.000 claims description 3
- 230000007850 degeneration Effects 0.000 claims description 3
- 238000004925 denaturation Methods 0.000 claims description 3
- 230000036425 denaturation Effects 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 3
- 238000002474 experimental method Methods 0.000 claims description 3
- 230000014759 maintenance of location Effects 0.000 claims description 3
- 238000009629 microbiological culture Methods 0.000 claims description 2
- 238000007865 diluting Methods 0.000 claims 2
- 235000011187 glycerol Nutrition 0.000 claims 2
- 241001434359 Muhlenbergia phleoides Species 0.000 claims 1
- 230000003321 amplification Effects 0.000 claims 1
- 238000003199 nucleic acid amplification method Methods 0.000 claims 1
- 230000001954 sterilising effect Effects 0.000 claims 1
- 244000005700 microbiome Species 0.000 abstract description 7
- 244000062720 Pennisetum compressum Species 0.000 description 6
- 239000000835 fiber Substances 0.000 description 6
- 235000019750 Crude protein Nutrition 0.000 description 5
- 244000130556 Pennisetum purpureum Species 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 244000005706 microflora Species 0.000 description 4
- 235000019082 Osmanthus Nutrition 0.000 description 3
- 241000333181 Osmanthus Species 0.000 description 3
- 238000004737 colorimetric analysis Methods 0.000 description 3
- 235000005911 diet Nutrition 0.000 description 3
- 230000037213 diet Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 150000007524 organic acids Chemical class 0.000 description 3
- 235000019629 palatability Nutrition 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 230000001953 sensory effect Effects 0.000 description 3
- OBMBUODDCOAJQP-UHFFFAOYSA-N 2-chloro-4-phenylquinoline Chemical compound C=12C=CC=CC2=NC(Cl)=CC=1C1=CC=CC=C1 OBMBUODDCOAJQP-UHFFFAOYSA-N 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 235000014633 carbohydrates Nutrition 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 239000003599 detergent Substances 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 235000012041 food component Nutrition 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 239000002932 luster Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 230000000050 nutritive effect Effects 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 239000012898 sample dilution Substances 0.000 description 2
- 239000003643 water by type Substances 0.000 description 2
- 241000305071 Enterobacterales Species 0.000 description 1
- 241001458359 Hemarthria compressa Species 0.000 description 1
- 241000282849 Ruminantia Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 1
- RJGDLRCDCYRQOQ-UHFFFAOYSA-N anthrone Chemical compound C1=CC=C2C(=O)C3=CC=CC=C3CC2=C1 RJGDLRCDCYRQOQ-UHFFFAOYSA-N 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- -1 benzene Phenol-hypochlorous acid Chemical compound 0.000 description 1
- 238000011325 biochemical measurement Methods 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 238000012875 competitive assay Methods 0.000 description 1
- 230000009514 concussion Effects 0.000 description 1
- 239000013256 coordination polymer Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 235000013325 dietary fiber Nutrition 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 244000144980 herd Species 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 238000000643 oven drying Methods 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 210000000697 sensory organ Anatomy 0.000 description 1
- 239000004460 silage Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000001117 sulphuric acid Substances 0.000 description 1
- 235000011149 sulphuric acid Nutrition 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
Classifications
-
- 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
- A23K30/00—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs
- A23K30/10—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder
- A23K30/15—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder using chemicals or microorganisms for ensilaging
- A23K30/18—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder using chemicals or microorganisms for ensilaging using microorganisms or enzymes
-
- 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
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Virology (AREA)
- Analytical Chemistry (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Animal Husbandry (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention discloses a kind of cibarium Wei Si Salmonellas and its application, screening calibration method, the cibarium Wei Si Salmonellas (Weissella cibaria) are preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is CGMCC No.15075；The cibarium Wei Si Salmonella growth rates are fast, and production acid is efficient；The cibarium Wei Si Salmonellas promote the quality of grass silage feed, are with a wide range of applications as grass silage additive.
Description
Technical field
The present invention relates to microbial technology fields, and in particular to a kind of cibarium Wei Si Salmonellas and its application, screening calibrating side
Method.
Background technology
Ensiling is the multiplication by lactic acid bacteria, and the fermentation substrate in raw material is changed into the acidic materials such as lactic acid, maintains acid
Property anaerobic environment, a kind of storage method preserved for a long time in favor of silo crop.Ensilage color is yellowish green, smell is sour fragrant, soft
Soft succulence, palatability are good, the extensive use in Animal husbandry production, especially in ruminant is raised, it has also become important is high-quality
Roughage source.
Lactic acid bacteria is the microoganism additives for ensiling promoted in recent years, and main function is micro- in adjusting ensiling material
Biotic component rapidly becomes dominant microflora, and the growth of Competitive assays harmful microorganism converts limited sugar, reduces ensiling pH value,
So as to improve the quality of ensilage, different its fermentation characters of ensiling material is in the presence of breasts that are different, and being suitable for ensiling at present
Sour bacterium single varieties, it is impossible to meet the market demand.
The content of the invention
In view of this, this application provides a kind of cibarium Wei Si Salmonellas, the cibarium Wei Si Salmonella growth rates are fast, production acid
It is efficient；For cibarium Wei Si Salmonellas as grass silage additive, fermented type is homofermentation, can quickly reduce grass silage pH
Value makes lactic acid bacteria become dominant microflora, accelerates fermenting-ripening, reduces the generation of ammoniacal nitrogen and add organic acid content, carry
The high fermentation quality of ensilage, so as to improve grass silage diet dry matter content, the dry matter rate of recovery, crude protein contain
Amount reduces fiber content, promotes grass silage quality of the fodder, is with a wide range of applications.
For solution more than technical problem, technical solution provided by the invention is a kind of cibarium Wei Si Salmonellas, the cibarium Wei
This Salmonella (Weissella cibaria) is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, ground
Location is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, deposit number CGMCC
No.15075, preservation date are on December 18th, 2017；
The 16s rDNA of above-mentioned cibarium Wei Si Salmonellas are as shown in SEQ ID NO.1.
The present invention also provides the above-mentioned applications for stating cibarium Wei Si Salmonellas in grass silage additive is prepared.
Preferably, the cibarium Wei Si Salmonellas are in Pennisetum ensiling and Hemarthria herbage additive is prepared
Using.
Preferably, the Pennisetum is that Pennisetum is that Gui Mu No.1 hybrid Chinese pennisetum, Gui Min draw napier grass.
Preferably, the Hemarthria herbage is Hemarthria compressa.
The present invention also provides the screening calibration methods of above-mentioned cibarium Wei Si Salmonellas, comprise the following steps：
1) Forage Grass is taken, is drawn after dilution and is applied to MRS solid medium cultures, chosen single bacterium colony and support and continue to cultivate
Afterwards, continuous passage culture is no less than 2 times, the bacterium colony purified；
2) bacterium colony for the purifying for obtaining step 1) is cultivated on lactic acid bacteria solid medium under the conditions of constant-temperatureanaerobic anaerobic
Afterwards, by Gram's staining and catalase haptoreaction, the identification of lactic acid bacteria is carried out, the lactic acid bacteria after identification is added in and is contained
It in the liquid nutrient media of sterile glycerol, and stores, the lactic acid bacteria isolated and purified；
3) after the lactic acid bacteria isolated and purified for obtaining step 2) carries out actication of culture, MRS Liquid Cultures are inoculated into
It is cultivated in base, obtains bacterium solution；Measure the bacterium solution pH and OD values, according to measurement result carry out screening pH=3.76, OD value=
1.641 bacterium solution obtains the bacterial strain after tentatively screening；
4) after the bacterial strain after the preliminary screening for obtaining step 3) carries out Anaerobic culturel, the DNA for carrying out single strain is carried
It takes, carrying out PCR amplification as template using the DNA of extraction obtains amplified production, and the amplified production is the cibarium Wei Si Salmonellas.
Preferably, the step specifically includes：
1) Forage Grass is taken, sterile distilled water is added in, is diluted to 10-1, 10-3With 10-5The sample dilution of three dilution factors
Liquid；Pipette samples dilution is applied to MRS solid medium cultures, continuous passage culture 2 times, the bacterium colony purified；
2) bacterium colony for the purifying for obtaining step 1) is on lactic acid bacteria solid medium under the conditions of 37 DEG C of constant-temperatureanaerobic anaerobics
After culture for 24 hours, by Gram's staining and catalase haptoreaction, the identification of lactic acid bacteria is carried out, by the lactic acid after identification
Bacterium is added in the liquid nutrient media containing sterile glycerol, and with -80 DEG C of storages, the lactic acid bacteria isolated and purified；
3) after the lactic acid bacteria isolated and purified for obtaining step 2) carries out actication of culture, it is inoculated with 3% inoculum concentration
Into 10ml MRS fluid nutrient mediums, with 40~45 DEG C of temperature, humidity 85%~98% cultivates 12h, obtains bacterium solution；Described in measure
Bacterium solution pH and OD values carry out the bacterium solution of screening pH=3.76, OD value=1.641 according to measurement result, after obtaining tentatively screening
Bacterial strain；
4) bacterial strain after the preliminary screening for obtaining step 3) carries out the Anaerobic culturel 48h under the conditions of 37 DEG C of temperature
Afterwards, the DNA extractions of single strain are carried out, carrying out PCR amplification as template using the DNA of extraction obtains amplified production, the amplified production
As described cibarium Wei Si Salmonellas.
Preferably, the DNA using extraction carries out PCR amplification as template and obtains amplified production process and be specially：With upstream
Primer：5 ,-AGAGTTTGATCCTGGCTCAG-3, anti-sense primer：5 ,-GGTTACCTTGTTACGACTT-3 are specific primer
PCR amplification is carried out for template to the DNA of extraction and obtains amplified production, the pcr amplification reaction condition is：95 DEG C of pre-degeneration 5min
Afterwards, 95 DEG C of denaturation 30s, 55 DEG C of annealing 30s, 72 DEG C of extension 90s are cycled 30 times, then 72 DEG C of extension 5min.
Preferably, the screening calibration method further includes：The cibarium Wei Si Salmonellas that step 4) is obtained pass through physiology
Biochemical measurement, Salt tolerance, different temperatures experiment, acid resistance test and sugar fermentating test evaluation test are analyzed.
Preferably, the screening calibration method further includes：The cibarium Wei Si Salmonellas that step 4) is obtained carry out 16S
RDNA sequence analyses.
Preferably, the Forage Grass is Pennisetum sample.
Preferably, the Pennisetum sample is Gui Mu No.1 hybrid Chinese pennisetum fresh grass, Gui Min draws napier grass fresh grass, osmanthus
It herds No. 1 hybrid Chinese pennisetum ensiling sample or napier grass ensiling sample is drawn in osmanthus Fujian.
Compared with prior art, detailed description are as follows by the application：
The present invention provides a kind of cibarium Wei Si Salmonellas, and growth rate is fast, and production acid is efficient.
Cibarium Wei Si Salmonellas provided by the invention are applied to the grass silage used time, can the limited sugar of rapid conversion, significantly
Ground reduces ensiling pH value, effectively improves the quality of ensilage, has larger potentiality and vast potential for future development.Further,
In herbage, the water-soluble carbohydrate content of Pennisetum is relatively low, and moisture is higher, and buffer index is larger,
Its stalk is hollow to have large quantity of air, so the fermentation of Chinese pennisetum ensiling is slow, ultimately results in ensiling failure.Hemarthria herbage is certainly
The lactic acid bacterium number of body attachment is less, and China at present reports Hemarthria Forage Research less.Cibarium Wei provided by the invention
For this Salmonella as grass silage additive, fermented type is heterofermentation, can quickly reduce grass silage pH value, make lactic acid bacteria into
For dominant microflora, accelerate fermenting-ripening, reduce the generation of ammonia nitrogen content and increase organic acid content, improve ensilage
Fermentation quality, so as to improve grass silage diet dry matter content, the dry matter rate of recovery, crude protein content promote grass silage
Quality of the fodder；Fiber content is reduced, improves palatability, and then promotes grass silage quality of the fodder, is had extensive
Application prospect.
The present invention provides Forage Grass in the screening calibration method of cibarium Wei Si Salmonellas through isolate and purify to obtain separate it is pure
The lactic acid bacteria of change, the lactic acid bacteria culturers activation culture from purifying, obtains bacterium solution；According to bacterium solution pH=3.76, OD value=1.641
Screening criteria is screened, and is obtained the bacterial strain after tentatively screening, after the bacterial strain after preliminary screening carries out Anaerobic culturel, is carried out single plant
The DNA extractions of bacterium carry out PCR amplification as template using the DNA of extraction and obtain amplified production, and method is simple and convenient, has extensive
Application prospect.
Screening calibration method the present invention provides cibarium Wei Si Salmonellas is come true by the identification to obtained microorganism
Surely the microorganism of institute's prescreening has been obtained as cibarium Wei Si Salmonellas provided by the invention, and repeatable implementation, widely should have
Use prospect.
Description of the drawings
It in order to illustrate more clearly about the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing
There is attached drawing needed in technology description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this
Some embodiments of invention, for those of ordinary skill in the art, without creative efforts, can be with
Other attached drawings are obtained according to these attached drawings.
Fig. 1 provides the optical microscope of cibarium Wei Si Salmonellas for the present invention.
Specific embodiment
In order to which those skilled in the art is made to more fully understand technical scheme, with reference to specific embodiment pair
The present invention is described in further detail.
A kind of cibarium Wei Si Salmonellas, the cibarium Wei Si Salmonellas (Weissella cibaria) are preserved in China Microbiological
Culture presevation administration committee common micro-organisms center, address are Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Chinese science
Institute of microbiology of institute, deposit number are CGMCC No.15075, and preservation date is on December 18th, 2017；
The 16s rDNA nucleotide of above-mentioned cibarium Wei Si Salmonellas (Weissella cibaria) is as shown in SEQ ID NO.1.
The screening calibration method of above-mentioned cibarium Wei Si Salmonellas, comprises the following steps：
1) Forage Grass is taken, sterile distilled water is added in, is diluted to 10-1, 10-3With 10-5The sample dilution of three dilution factors
Liquid；Pipette samples dilution is applied to MRS solid medium cultures, continuous passage culture 2 times, the bacterium colony purified.
2) bacterium colony for the purifying for obtaining step 1) is on lactic acid bacteria solid medium under the conditions of 37 DEG C of constant-temperatureanaerobic anaerobics
After culture for 24 hours, by Gram's staining and catalase haptoreaction, the identification of lactic acid bacteria is carried out, by the lactic acid after identification
Bacterium is added in the liquid nutrient media containing sterile glycerol, and with -80 DEG C of storages, the lactic acid bacteria isolated and purified.
3) after the lactic acid bacteria isolated and purified for obtaining step 2) carries out actication of culture, it is inoculated with 3% inoculum concentration
Into 10ml MRS fluid nutrient mediums, with 40~45 DEG C of temperature, humidity 85%~98% cultivates 12h, obtains bacterium solution；Described in measure
Bacterium solution pH and OD values, measurement result are shown in Table 1, the bacterium solution of screening pH=3.76, OD value=1.641 are carried out according to measurement result, is obtained
Bacterial strain to after preliminary screening.
4) bacterial strain after the preliminary screening for obtaining step 3) carries out the Anaerobic culturel 48h under the conditions of 37 DEG C of temperature
Afterwards, the DNA extractions of single strain are carried out, with sense primer：27F (5 ,-AGAGTTTGATCCTGGCTCAG-3), anti-sense primer：
1492R (5 ,-GGTTACCTTGTTACGACTT-3) is that specific primer is that template progress PCR amplification is obtained to the DNA of extraction
Amplified production, the amplified production are above-mentioned cibarium Wei Si Salmonellas；The pcr amplification reaction condition is：95 DEG C of pre-degenerations
After 5min, 95 DEG C of denaturation 30s, 55 DEG C of annealing 30s, 72 DEG C of extension 90s are cycled 30 times, then 72 DEG C of extension 5min；
Wherein, the Forage Grass is Pennisetum sample；The Pennisetum sample hybridizes for Gui Mu No.1
Chinese pennisetum fresh grass, Gui Min draw napier grass fresh grass, Gui Mu No.1 hybrid Chinese pennisetum ensiling sample or osmanthus Fujian and draw napier grass ensiling sample.
Fig. 1 is the above-mentioned optical microscope for providing cibarium Wei Si Salmonellas.
1 bacterium solution pH of table and OD values
The microorganism of strain number F10 is the bacterial strain after this patent tentatively screening, by the bacterial strain after the preliminary screening through detesting
Oxygen culture, DNA extractions, the cibarium Wei Si Salmonellas provided by the invention obtained using the DNA of extraction as template progress PCR amplification,
Growth rate is fast, and production acid is efficient.
Embodiment 2
Physiological and biochemical test：The cibarium Wei Si Salmonella physiological and biochemical properties in embodiment 1 are analyzed, the results are shown in Table 2.
Different temperatures is tested：Cibarium Wei Si Salmonellas in embodiment 1 are inoculated into MRS Liquid Cultures with 3% inoculum concentration
It in base, is cultivated under the temperature conditionss of table 2, observes its growth situation, the results are shown in Table 2.
Salt tolerance：By the cibarium Wei Si Salmonellas in embodiment 1 to contain NaCl concentration 3.0% (w/v) and 6.5% (w/
V) in MRS fluid nutrient mediums, after being cultivated 2 days at 30 DEG C, its salt tolerance is measured, the results are shown in Table 2.
Acid resistance test：It it is 30 DEG C in the MRS fluid nutrient mediums of table 2pH value conditions by the cibarium Wei Si Salmonellas in embodiment 1
After lower culture 7 days, its acid resistance is measured, the results are shown in Table 2.
Sugar fermentating test：Cibarium Wei Si Salmonella sugar fermenting characteristics in embodiment 1 are analyzed by sugar fermentating test, the result is shown in
Table 3.
16S rDNA sequence analyses：Cibarium Wei Si Salmonellas in embodiment 1 are subjected to 16S rDNA sequence analyses, 16s
RDNA is as shown in SEQ ID NO.1；With other cibariums Wei Si Salmonellas (Weissella cibaria) 16S rDNA analytical sequences
Similitude reaches 100% and 99%.
2 physiological and biochemical test of table, different temperatures experiment, Salt tolerance and acid resistance test result
In table 2 ,+：Grow Positive；-:Do not grow Negative；W:Faint growth Weakly positive.
As shown in Table 2, cibarium Wei Si Salmonellas salt tolerance and acid resistance provided by the invention are good, to the adaptability of different temperatures
Preferably.
3 sugar fermentating test result of table
In table 3 ,+：Grow Positive；-:Do not grow Negative；W:Faint growth Weakly positive.
As shown in Table 3, cibarium Wei Si Salmonellas provided by the invention are handed over wide using carbon source scope, can preferably utilize different green grass or young crops
Different types of carbon source is fermented in storage raw material.
Embodiment 3
Using the cibarium Wei Si Salmonellas of the present invention as grass silage additive
The herbage of 1.5~2m of height is taken in Chongzhou City of Sichuan Agricultural University base, it is about 20mm to be cut into length, obtains fresh sample,
Each processing respectively takes fresh sample 300g to put into polybag (25 × 35cm；Aodeju, China), according to by the present invention in embodiment 1
Cibarium Wei Si Salmonellas 1.0 × 108CFU/g fresh materials add 1ml in every bag of ensilage, and then vacuum-pumping and sealing, is placed in
It ferments in room temperature, each 3 repetitions of processing group break a seal after ensiling 60d, obtain ensiling sample.
1st, subjective appreciation：
1) the sense organ ensiling standards of grading and ranking pair that ensiling sample is compiled and edit according to German animal husbandry association (DLG) for 1899
Smell, structure and the color and luster of ensilage carry out sensory evaluation scores respectively.
4 ensilage subjective appreciation standard (DLG) of table
2) results of sensory evaluation is shown in Table 5.
5 results of sensory evaluation of table
As shown in Table 5, cibarium Wei Si Salmonellas provided by the invention do not add as grass silage additive and any addition
The control group of agent is compared, and in terms of smell, stink etc. is weak, and more preferably, structure of stem and leaf is kept more preferably armaticity, and color and luster is more preferable.
2nd, fermentation quality is analyzed：
1) 20g ensilings sample addition 180mL deionized waters is taken to be put into pony mixer and stir 1min, 8 layers of filter cloth is crossed, uses benzene
Phenol-hypochlorous acid receives colorimetric method for determining filtrate pH value, and colorimetric method for determining filtrate ammoniacal nitrogen (NH is received with phenol-hypochlorous acid3-N)
20g ensilings sample is taken to mix concussion with 180ml sterile purified waters, with four layers of filtered through gauze, using high performance liquid chromatography
Method measures organic acid content.
2) herbage be Pennisetum, Pennisetum fermentation quality analysis in table 6, table 7.
6 Pennisetum fermentation quality of table is analyzed
7 Pennisetum fermentation quality of table is analyzed
ND:It does not detect.
3) herbage is Hemarthria herbage, and Hemarthria herbage fermentation quality analysis result is shown in Table 8.
8 Hemarthria herbage fermentation quality of table is analyzed
From table 6~8, cibarium Wei Si Salmonellas provided by the invention are as grass silage additive with not adding any add
The control group of agent is added to compare, can quickly reduce grass silage pH value, Silage Quality is more preferable, cibarium Wei Si Salmonellas provided by the invention
As grass silage additive NH3- N/%TN does not add the control group of any additive substantially less than.Cibarium provided by the invention
Wei Si Salmonellas can dramatically increase lactic acid content as grass silage additive, can promote lactic acid bacterium number, and significantly reduce yeast
Bacterium and enterobacteria quantity are that lactic acid bacteria becomes dominant microflora, always the harmful bacteria in ensilage, improve the ensiling product of herbage
Matter.Cibarium Wei Si Salmonellas provided by the invention are remarkably improved organic as Pennisetum additives for ensiling, ensiling 60d
Acid content improves the fermentation rate of Pennisetum ensiling, so as to improve ensiling success rate, improves the green grass or young crops of Pennisetum
Store quality.
3rd, Analysis of Nutritive Composition：
1) ensiling sample is taken to be placed in air dry oven, 105 DEG C of water-removing 0.5h then dry 72h or so until constant weight at 65 DEG C,
Drying sample smashes it through 40 mesh sieves with small plant sample pulverizer.Using conventional oven drying method measure dry matter (Dry Matter,
DM), using 8400 type full-automatic Kjeldahl determination device determination of distillation crude protein (Crude Protein, CP) of FOSS, using Fan Shi
It washs fibre method and measures neutral detergent fiber (Neutral Detergent Fiber, NDF) and acid detergent fiber (Acid
Detergent Fiber, ADF), using Anthrone Sulphuric acid Colorimetry soluble-carbohydrate (Water Soluble
Carbohydrates,WSC)；
2) herbage is Pennisetum, and Pennisetum Analysis of Nutritive Composition the results are shown in Table 9.
9 Pennisetum nutritional ingredient of table
3) herbage is Hemarthria herbage, and Hemarthria nutritional components the results are shown in Table 10.
10 Hemarthria nutritional components of table
From table 9, table 10, cibarium Wei Si Salmonellas provided by the invention are any with not adding as grass silage additive
The control group of additive is compared, and can improve grass silage diet dry matter content, and the dry matter rate of recovery, crude protein content are promoted
Grass silage quality of the fodder；Fiber content is reduced, palatability is improved, and then promotes grass silage quality of the fodder.
It the above is only the preferred embodiment of the present invention, it is noted that above-mentioned preferred embodiment is not construed as pair
The limitation of the present invention, protection scope of the present invention should be subject to claim limited range.For the art
For those of ordinary skill, without departing from the spirit and scope of the present invention, several improvements and modifications can also be made, these change
Protection scope of the present invention is also should be regarded as into retouching.
Sequence table
<110>Sichuan Agricultural University
<120>A kind of cibarium Wei Si Salmonellas and its application, screening calibration method
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1390
<212> DNA
<213>Cibarium Wei Si Salmonellas (Weissella cibaria)
<400> 1
ggctttgggt gttacaaact ctcatggtgt gacgggcggt gtgtacaaga cccgggaacg 60
tattcaccgc ggcgtgctga tccgcgatta ctagcgattc cgacttcatg taggcgagtt 120
gcagcctaca atccgaactg agacgtactt taagagatta gctcaccctc gcgggttggc 180
aactcgttgt atacgccatt gtagcacgtg tgtagcccag gtcataaggg gcatgatgat 240
ttgacgtcat ccccaccttc ctccggtttg tcaccggcag tctcactaga gtgcccaact 300
aaatgctggc aactagtaat aagggttgcg ctcgttgcgg gacttaaccc aacatctcac 360
gacacgagct gacgacaacc atgcaccacc tgtcaccttg tccccgaagg gaacgctcca 420
tctctggagt tgtcaaggga tgtcaagacc tggtaaggtt cttcgcgttg cttcgaatta 480
aaccacatgc tccaccgctt gtgcgggtcc ccgtcaattc ctttgagttt caaccttgcg 540
gtcgtactcc ccaggcggag tgcttaatgc gttagctgcg gcacttaagg gcggaaaccc 600
tcaaacacct agcactcatc gtttacggtg tggactacca gggtatctaa tcctgtttgc 660
tacccacact ttcgagcctc aacgtcagtt acagtccaga aagccgcctt cgccactggt 720
gttcttccat atatctacgc atttcaccgc tacacatgga gttccacttt cctctactgc 780
actcaagtca tccagtttcc aaagcaattc ctcagttgag ctgagggctt tcacttcaga 840
cttaaataac cgtctgcgct cgctttacgc ccaataaatc cggataacgc ttggaacata 900
cgtattaccg cggctgctgg cacgtattta gccgttcctt tctggtaaga taccgtcaca 960
cattgaacag ttactctcaa tgtcattctt ctcttacaac agtgttttac gagccgaaac 1020
ccttcatcac acacgcggcg ttgctccatc aggctttcgc ccattgtgga agattcccta 1080
ctgctgcctc ccgtaggagt atgggccgtg tctcagtccc attgtggccg atcagtctct 1140
caactcggct atgcatcatc gtcttggtga gccattacct caccaactaa ctaatgcacc 1200
gcgggaccat ctcttagtga tagcagaacc atcttttaag tagcaaccat gcggttgcta 1260
ttgttatacg gtattagcat ctgtttccaa atgttatccc ctgctaagag gtaggtttcc 1320
cacgtgttac tcacccgttc gccactcttt gcaatgtcca tcgtcatatc tgagcaagct 1380
cttcaaatca 1390
<210> 2
<211> 20
<212> DNA
<213>Artificial sequence (unknown)
<400> 2
agagtttgat cctggctcag 20
<210> 3
<211> 19
<212> DNA
<213>Artificial sequence (unknown)
<400> 3
ggttaccttg ttacgactt 19
Claims (10)
1. a kind of cibarium Wei Si Salmonellas, which is characterized in that during the cibarium Wei Si Salmonellas (Weissella cibaria) are preserved in
State's Microbiological Culture Collection administration committee common micro-organisms center, deposit number are CGMCC No.15075.
2. cibarium Wei Si Salmonellas described in claim 1, which is characterized in that the 16srDNA such as SEQ of the cibarium Wei Si Salmonellas
Shown in ID NO.1.
3. the application of cibarium Wei Si Salmonellas described in claim 1, which is characterized in that the cibarium Wei Si Salmonellas are preparing herbage
Application in additives for ensiling.
4. the application of the cibarium Wei Si Salmonellas described in claim 3, which is characterized in that the cibarium Wei Si Salmonellas are preparing wolf tail
Grass belongs to the application in herbage and Hemarthria grass silage additive.
5. a kind of screening calibration method of cibarium Wei Si Salmonellas described in claim 1, which is characterized in that comprise the following steps：
1) Forage Grass is taken, is drawn after dilution and is applied to MRS solid medium cultures, is chosen after single bacterium colony supports and continue culture, even
Continuous secondary culture is no less than 2 times, the bacterium colony purified；
2) after the bacterium colony for the purifying for obtaining step 1) is cultivated on lactic acid bacteria solid medium under the conditions of constant-temperatureanaerobic anaerobic, lead to
Gram's staining and catalase haptoreaction are crossed, carries out the identification of lactic acid bacteria, the lactic acid bacteria after identification is added in containing sterilizing
It in the liquid nutrient media of glycerine, and stores, the lactic acid bacteria isolated and purified；
3) after the lactic acid bacteria isolated and purified for obtaining step 2) carries out actication of culture, it is inoculated into MRS fluid nutrient mediums
Culture, obtains bacterium solution；The bacterium solution pH and OD values are measured, screening pH=3.76, OD value=1.641 are carried out according to measurement result
Bacterium solution obtains the bacterial strain after tentatively screening；
4) after the bacterial strain after the preliminary screening for obtaining step 3) carries out Anaerobic culturel, the DNA extractions of single strain are carried out, with
The DNA of extraction carries out PCR amplification for template and obtains amplified production, and the amplified production is the cibarium Wei Si Salmonellas.
6. screening calibration method according to claim 5, which is characterized in that the Forage Grass is Pennisetum sample
Product.
7. screening calibration method according to claim 5, which is characterized in that the step specifically includes：
1) Forage Grass is taken, sterile distilled water is added in, is diluted to 10-1, 10-3With 10-5The sample diluting liquid of three dilution factors；It inhales
Sample diluting liquid is taken to be applied to MRS solid medium cultures, continuous passage culture 2 times, the bacterium colony purified；
2) bacterium colony for the purifying for obtaining step 1) is cultivated on lactic acid bacteria solid medium under the conditions of 37 DEG C of constant-temperatureanaerobic anaerobics
After for 24 hours, by Gram's staining and catalase haptoreaction, the identification of lactic acid bacteria is carried out, the lactic acid bacteria after identification is added
Enter in the liquid nutrient media containing sterile glycerol, and with -80 DEG C of storages, the lactic acid bacteria isolated and purified；
3) after the lactic acid bacteria isolated and purified for obtaining step 2) carries out actication of culture, it is inoculated into 3% inoculum concentration
In 10ml MRS fluid nutrient mediums, with 40~45 DEG C of temperature, humidity 85%~98% cultivates 12h, obtains bacterium solution；Measure the bacterium
Liquid pH and OD values carry out the bacterium solution of screening pH=3.76, OD value=1.641 according to measurement result, obtain the bacterium after tentatively screening
Strain；
4) bacterial strain after the preliminary screening for obtaining step 3) is carried out after Anaerobic culturel 48h under the conditions of 37 DEG C of temperature, into
The DNA extractions of row single strain carry out PCR amplification as template using the DNA of extraction and obtain amplified production, and the amplified production is institute
State cibarium Wei Si Salmonellas.
8. screening calibration method according to claim 5, which is characterized in that the DNA using extraction carries out PCR as template
Amplification obtains amplified production process and is specially：With sense primer：5 ,-AGAGTTTGATCCTGGCTCAG-3, anti-sense primer：5 ,-
GGTTACCTTGTTACGACTT-3 is that specific primer is that template progress PCR amplification obtains amplified production to the DNA of extraction, institute
Stating pcr amplification reaction condition is：After 95 DEG C of pre-degeneration 5min, 95 DEG C of denaturation 30s, 55 DEG C of annealing 30s, 72 DEG C extend 90s, cycle
30 times, then 72 DEG C of extension 5min.
9. screening calibration method according to claim 5, which is characterized in that the screening calibration method further includes：It will step
The rapid cibarium Wei Si Salmonellas 4) obtained pass through physiological and biochemical test, Salt tolerance, different temperatures experiment, acid resistance test and sugar
Fermentation test evaluation test is analyzed.
10. screening calibration method according to claim 5, which is characterized in that the screening calibration method further includes：It will step
The rapid cibarium Wei Si Salmonellas 4) obtained carry out 16S rDNA sequence analyses.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810093014.0A CN108060102B (en) | 2018-01-29 | 2018-01-29 | A kind of cibarium Wei Si Salmonella and its application, screening calibration method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810093014.0A CN108060102B (en) | 2018-01-29 | 2018-01-29 | A kind of cibarium Wei Si Salmonella and its application, screening calibration method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108060102A true CN108060102A (en) | 2018-05-22 |
| CN108060102B CN108060102B (en) | 2019-01-04 |
Family
ID=62134318
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810093014.0A Active CN108060102B (en) | 2018-01-29 | 2018-01-29 | A kind of cibarium Wei Si Salmonella and its application, screening calibration method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108060102B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109402010A (en) * | 2018-11-16 | 2019-03-01 | 大连工业大学 | A method of utilizing Lactococcus lactis M10 and the smelly mandarin fish of cibarium Wei Si Salmonella M3 combined inoculation Rapid Fermentation |
| CN114703084A (en) * | 2022-01-28 | 2022-07-05 | 黑龙江八一农垦大学 | Weissella cibaria with broad-spectrum antibacterial activity |
| CN117229980A (en) * | 2023-11-08 | 2023-12-15 | 北京市农林科学院 | Weissella sinica fermented feed and application and deodorization effect thereof |
-
2018
- 2018-01-29 CN CN201810093014.0A patent/CN108060102B/en active Active
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109402010A (en) * | 2018-11-16 | 2019-03-01 | 大连工业大学 | A method of utilizing Lactococcus lactis M10 and the smelly mandarin fish of cibarium Wei Si Salmonella M3 combined inoculation Rapid Fermentation |
| CN109402010B (en) * | 2018-11-16 | 2021-09-24 | 大连工业大学 | Method for rapidly fermenting smelly mandarin fish by mixed inoculation of lactococcus lactis M10 and Weissella cibaria M3 |
| CN114703084A (en) * | 2022-01-28 | 2022-07-05 | 黑龙江八一农垦大学 | Weissella cibaria with broad-spectrum antibacterial activity |
| CN114703084B (en) * | 2022-01-28 | 2023-06-23 | 黑龙江八一农垦大学 | Weissella food with broad-spectrum antibacterial activity |
| CN117229980A (en) * | 2023-11-08 | 2023-12-15 | 北京市农林科学院 | Weissella sinica fermented feed and application and deodorization effect thereof |
| CN117229980B (en) * | 2023-11-08 | 2024-01-30 | 北京市农林科学院 | Weissella sinica fermented feed and application and deodorization effect thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108060102B (en) | 2019-01-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108060104B (en) | A kind of lactobacillus plantarum and its application, screening calibration method | |
| CN107047978A (en) | A kind of Lactobacillus plantarum and its application in ensilage is prepared | |
| CN107034156A (en) | Lactobacillus plantarum and its application | |
| CN102640885B (en) | Applications of Lactobacillus plantarum in complete-strain Triticumspp silage | |
| CN108060102B (en) | A kind of cibarium Wei Si Salmonella and its application, screening calibration method | |
| CN108060103B (en) | A kind of Pediococcus pentosaceus and its application, screening calibration method | |
| CN108265016B (en) | A kind of lactic bacteria additive and its application, preparation method | |
| CN111269849B (en) | Lactobacillus plantarum and application thereof | |
| CN111304112B (en) | Composite microbial inoculum for ensiling sugarcane tail leaves | |
| CN113308418A (en) | Lactobacillus chaff for fermentation and fermentation preparation process thereof | |
| CN114908017B (en) | Lactobacillus plantarum and application thereof in silage | |
| CN107653203A (en) | A kind of Lactobacillus rhamnosus and its application in feed addictive is prepared | |
| CN115011515B (en) | Strain for increasing plant alcohol content in alfalfa silage and application thereof | |
| CN111748490A (en) | Lactobacillus sake and application thereof | |
| CN114908018B (en) | Lactobacillus brevis and application thereof in silage | |
| CN115161234B (en) | Bacillus bailii and application thereof in silage preservation | |
| CN114181867B (en) | Lactobacillus plantarum, and product and application thereof | |
| CN113897312B (en) | Preparation and application of animal feeding microbial inoculum | |
| CN115702668B (en) | Mixed silage method of cyperus esculentus stems and leaves and rape straw | |
| CN106615609B (en) | Lactobacillus buchneri capable of improving silage quality and application thereof | |
| CN111040969B (en) | Compound lactobacillus agent and application thereof in buffalo silage | |
| CN114231464A (en) | Bacillus coagulans and application thereof | |
| CN114276949A (en) | Lactobacillus paracasei and application thereof in preparation of broussonetia papyrifera silage | |
| CN112715758A (en) | Lactobacillus buchneri BL4 and method for preparing silage by using same | |
| CN109971667A (en) | One plant of porcine lactobacillus plantarum and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |