CN105483050B - One lactobacillus plantarum and its application in fermented feed - Google Patents
One lactobacillus plantarum and its application in fermented feed Download PDFInfo
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- 240000006024 Lactobacillus plantarum Species 0.000 title claims abstract description 38
- 235000013965 Lactobacillus plantarum Nutrition 0.000 title claims description 31
- 229940072205 lactobacillus plantarum Drugs 0.000 title claims description 31
- 244000005700 microbiome Species 0.000 claims abstract description 12
- 238000004321 preservation Methods 0.000 claims abstract description 10
- 230000001580 bacterial effect Effects 0.000 claims abstract description 5
- 238000000855 fermentation Methods 0.000 claims description 35
- 230000004151 fermentation Effects 0.000 claims description 35
- 235000013312 flour Nutrition 0.000 claims description 30
- 239000000843 powder Substances 0.000 claims description 21
- 235000007164 Oryza sativa Nutrition 0.000 claims description 18
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 18
- 235000009566 rice Nutrition 0.000 claims description 18
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 17
- 239000007788 liquid Substances 0.000 claims description 17
- 239000000463 material Substances 0.000 claims description 15
- 244000068988 Glycine max Species 0.000 claims description 12
- 235000010469 Glycine max Nutrition 0.000 claims description 12
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 12
- 239000000203 mixture Substances 0.000 claims description 12
- 235000013339 cereals Nutrition 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- 240000008042 Zea mays Species 0.000 claims description 8
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 8
- 235000005822 corn Nutrition 0.000 claims description 8
- 239000002054 inoculum Substances 0.000 claims description 7
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 5
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims description 5
- 241000235342 Saccharomycetes Species 0.000 claims description 5
- 241000209140 Triticum Species 0.000 claims description 5
- 235000021307 Triticum Nutrition 0.000 claims description 5
- 229940088594 vitamin Drugs 0.000 claims description 5
- 235000013343 vitamin Nutrition 0.000 claims description 5
- 239000011782 vitamin Substances 0.000 claims description 5
- 229930003231 vitamin Natural products 0.000 claims description 5
- 235000019733 Fish meal Nutrition 0.000 claims description 4
- 244000017020 Ipomoea batatas Species 0.000 claims description 4
- 235000002678 Ipomoea batatas Nutrition 0.000 claims description 4
- 235000019764 Soybean Meal Nutrition 0.000 claims description 4
- 241000482268 Zea mays subsp. mays Species 0.000 claims description 4
- 150000001413 amino acids Chemical class 0.000 claims description 4
- 239000004467 fishmeal Substances 0.000 claims description 4
- 235000021552 granulated sugar Nutrition 0.000 claims description 4
- 150000003839 salts Chemical class 0.000 claims description 4
- 239000004455 soybean meal Substances 0.000 claims description 4
- 150000003722 vitamin derivatives Chemical class 0.000 claims description 4
- YYRMJZQKEFZXMX-UHFFFAOYSA-L calcium bis(dihydrogenphosphate) Chemical compound [Ca+2].OP(O)([O-])=O.OP(O)([O-])=O YYRMJZQKEFZXMX-UHFFFAOYSA-L 0.000 claims description 3
- 229940062672 calcium dihydrogen phosphate Drugs 0.000 claims description 3
- 229910000389 calcium phosphate Inorganic materials 0.000 claims description 3
- 235000019691 monocalcium phosphate Nutrition 0.000 claims description 3
- -1 microelement Chemical class 0.000 claims description 2
- 240000007594 Oryza sativa Species 0.000 claims 7
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 abstract description 18
- 239000002253 acid Substances 0.000 abstract description 10
- 239000004310 lactic acid Substances 0.000 abstract description 9
- 235000014655 lactic acid Nutrition 0.000 abstract description 9
- 201000010099 disease Diseases 0.000 abstract description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 5
- 230000036039 immunity Effects 0.000 abstract description 5
- 230000003031 feeding effect Effects 0.000 abstract description 4
- 210000000936 intestine Anatomy 0.000 abstract description 4
- 244000144972 livestock Species 0.000 abstract description 4
- 235000013336 milk Nutrition 0.000 abstract description 4
- 239000008267 milk Substances 0.000 abstract description 4
- 210000004080 milk Anatomy 0.000 abstract description 4
- 244000144977 poultry Species 0.000 abstract description 4
- 244000052616 bacterial pathogen Species 0.000 abstract description 3
- 235000019621 digestibility Nutrition 0.000 abstract description 3
- 241000186660 Lactobacillus Species 0.000 description 13
- 229940039696 lactobacillus Drugs 0.000 description 13
- 239000002609 medium Substances 0.000 description 13
- 241000209094 Oryza Species 0.000 description 11
- 239000000243 solution Substances 0.000 description 8
- 230000001954 sterilising effect Effects 0.000 description 8
- 238000002474 experimental method Methods 0.000 description 7
- 239000001963 growth medium Substances 0.000 description 7
- 238000002360 preparation method Methods 0.000 description 6
- 206010012735 Diarrhoea Diseases 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 4
- 238000002835 absorbance Methods 0.000 description 4
- 239000005030 aluminium foil Substances 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 239000012530 fluid Substances 0.000 description 4
- 238000009629 microbiological culture Methods 0.000 description 4
- 235000015097 nutrients Nutrition 0.000 description 4
- 108020004465 16S ribosomal RNA Proteins 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 235000012054 meals Nutrition 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 2
- 101000869690 Homo sapiens Protein S100-A8 Proteins 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 102100032442 Protein S100-A8 Human genes 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 229960005069 calcium Drugs 0.000 description 2
- 239000011575 calcium Substances 0.000 description 2
- 229910052791 calcium Inorganic materials 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 201000010538 Lactose Intolerance Diseases 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 241000344273 bacterium 36 Species 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- UUVBYOGFRMMMQL-UHFFFAOYSA-N calcium;phosphoric acid Chemical compound [Ca].OP(O)(O)=O UUVBYOGFRMMMQL-UHFFFAOYSA-N 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 230000034303 cell budding Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 235000020247 cow milk Nutrition 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000003640 drug residue Substances 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 239000011344 liquid material Substances 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 229940099596 manganese sulfate Drugs 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000006872 mrs medium Substances 0.000 description 1
- 235000019629 palatability Nutrition 0.000 description 1
- 238000009928 pasteurization Methods 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 101150080777 pheS gene Proteins 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000011218 seed culture Methods 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 1
- 229940038773 trisodium citrate Drugs 0.000 description 1
- 230000004584 weight gain Effects 0.000 description 1
- 235000019786 weight gain Nutrition 0.000 description 1
- 239000012138 yeast extract Substances 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
-
- 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
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Fodder In General (AREA)
Abstract
The invention discloses a lactobacillus plantarum Lactobacillus plantarum, it is characterized in that, the bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation place is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica, the number that preservation is registered on the books is CGMCC No.11721, and the deposit date is on November 23rd, 2015.Strain provided by the invention growth, produce acid, in terms of show good performance, fermented product has good feeding effect.It, which is metabolized lactic acid, can inhibit enteric pathogenic bacteria growth, so as to be obviously improved the immunity of livestock and poultry, improves intestine microenvironment, reduces disease incidence, and bring certain promotion to digestibility, the output of milk etc..
Description
Technical field
The present invention relates to microbe applications and feed preparation manufacture field, more particularly to one kind for promoting animal immune
The lactobacillus plantarum of power and its application in fermented feed.
Background technique
Fermented feed have palatability it is good, safe and pollution-free, drug residue free, it is easy to digest the features such as.It is fermented by feeding
Feed can promote animal immunizing power, reduce the generation of disease, reduce diarrhea rate, death rate etc., it is possible to reduce or substitution antibiosis
The use of element.
Lactobacillus plantarum fermentation process can use the lactose in raw material and generate lactic acid, and lactic acid can promote calcium in conjunction with calcium
Absorption, therefore the group for there is lactose intolerance is particularly in and quickly generates budding group, and fermented feed has very big
Advantage.Feed produces various lactobacillus metabolite by lactobacillus-fermented, including vitamins, enzyme and various biologies are lived
Property substance etc., fermented feed is easier to digest and assimilate.But the fermentability of current lactobacillus plantarum is limited, raises in fermentation
Production application in material is ineffective.
Summary of the invention
In view of this, it is an object of the invention to propose a lactobacillus plantarum and its application in fermented feed, with
The immunity for promoting animal reduces the generation of disease, reduces diarrhea rate, death rate etc..
Based on above-mentioned purpose, lactobacillus plantarum Lactobacillus plantarum provided by the invention is preserved in China
Microbiological Culture Collection administration committee common micro-organisms center, preservation place are Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Institute of Microorganism, Academia Sinica, the number that preservation is registered on the books are CGMCC No.11721, and the deposit date is 2015 11
The moon 23.
The present invention also provides a kind of above-mentioned object lactobacillus Lactobacillus plantarum answering in fermented feed
With.
In some embodiments of the invention, the application includes:
Into cereal materials plus water mixes, and fermentation medium is made, by the lactobacillus plantarum Lactobacillus
Plantarum, which is inoculated in the fermentation medium, to ferment, and fermentation liquid is made；
Non-fermented raw mixture is mixed with fermentation liquid, fermented feed is made.
In some embodiments of the invention, by the lactobacillus plantarum Lactobacillus plantarum and wine brewing
Yeast Saccharomyces cerevisiae, which is inoculated in the fermentation medium, carries out cooperative fermentation.
In some embodiments of the invention, the saccharomycete is Saccharomyces Cerevisiae in S accharomyces cerevisiae
Sa-10 has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, CGMCC No.6120.
In some embodiments of the invention, the cereal materials are popcorn, dehulled soybean meal or fermented bean dregs, sweet potato
Powder or soybean powder, rice flour or small rice and flour, wheat flour, the popcorn, dehulled soybean meal or fermented bean dregs, sweet potato powder or Huang
The weight percentage of bean powder, rice flour or small rice and flour, wheat flour and water is followed successively by 1~20%, 1~50%, 1~20%, 1
~20%, 1~20% and 50~90%.
In some embodiments of the invention, the cereal materials are rice flour, soybean powder and corn flour, the rice
The weight percentage in face, soybean powder and corn flour is followed successively by 1~30%, 1~30% and 40~80%.
In some embodiments of the invention, the inoculum concentration of the lactobacillus plantarum Lactobacillus plantarum
It is 1%~3%.
In some embodiments of the invention, the non-fermented raw mixture includes amino acid, vitamin, micro member
Element, salt, mountain flour, calcium dihydrogen phosphate, white granulated sugar and super steam fish meal.
Strain provided by the invention growth, produce acid, in terms of show good performance, fermented product has
Good feeding effect.It, which is metabolized lactic acid, can inhibit enteric pathogenic bacteria growth, so as to be obviously improved the immunity of livestock and poultry, change
Kind intestine microenvironment, reduces disease incidence, and bring certain promotion to digestibility, the output of milk etc..
Specific embodiment
To make the objectives, technical solutions, and advantages of the present invention clearer, below in conjunction with specific embodiment, to this hair
Bright further description.
The separation and identification of 1 lactobacillus plantarum Lactobacillus plantarum of embodiment
It is described to state lactobacillus plantarum EM-12 bacterial strain and be deposited in Chinese microorganism strain preservation management on November 23rd, 2015
Committee's common micro-organisms center, deposit number are as follows: CGMCC No.11721, depositary institution address: the Chaoyang District, Beijing City North Star
The institute 3 of West Road 1, Institute of Microorganism, Academia Sinica, postcode: 100101.
Lactobacillus plantarum EM-12 provided by the invention is to make bolter in fermentation cow's milk by oneself from four prince flag herdsman of the Inner Mongol
Choosing obtains, and specific separation authentication step is as follows:
1) it takes rice meal, soybean powder and corn flour to mix by weight the ratio of 1:1:1, takes said mixture according to material water
The ratio of weight ratio 1:4 adds water to mix, and culture medium is made；
2) the fermentation milk sample containing viable bacteria is taken, is inoculated in the culture medium according to 2% inoculum concentration, is fullyd shake mixed
It is even, 36 DEG C of anaerobism, stationary cultures, incubation time 24 hours；Above-mentioned culture solution is passed on 3 times repeatedly in this culture medium, is obtained
It is suitble to the lactobacillus strain of fermented cereal class；
3) above-mentioned culture solution 1mL is taken, gradient dilution is inoculated with and is coated on 36 DEG C of culture 72h on MRS plate；Picking individual colonies
Bacterium 36 DEG C of culture 18h in MRS fluid nutrient medium are met, in case preservation and identification use.
4) strain is sent to Institute of Microorganism, Academia Sinica and is identified, according to bacterium cell form, biochemical character,
The comprehensive analysis such as 16S rRNA gene order, pheS gene order, are accredited as lactobacillus plantarum (Lactobacillus
plantarum)。
The cellular morphology and physical and chemical experiment result of the bacterial strain see the table below:
16S rDNA sequencing results:
GCGTGCTAATACATGCAGTCGAACGAACTCTGGTATTGATTGGTGCTTGCATCATGATTTACATTTGAG
TGAGTGGCGAACTGGTGAGTAACACGTGGGAAACCTGCCCAGAAGCGGGGGATAACACCTGGAAACAGATGCTAATA
CCGCATAACAACTTGGACCGCATGGTCCGAGTTTGAAAGATGGCTTCGGCTATCACTTTTGGATGGTCCCGCGGCGT
ATTAGCTAGATGGTGGGGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGAGAGGGTAATCGGCCACATTGG
GACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCA
ACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAACATATCTGAGAGTAACTGTTC
AGGTATTGACGGTATTTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCG
TTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCTTCGGCTCAACCGAA
GAAGTGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGA
TATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGTATGGGTAGCAA
ACAGGATTAGATACCCTGGTAGTCCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCT
GCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGGCCGCAAGGCTGAAACTCAAAGGAATTGACGGGGGCCCGC
ACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATACTATGCAAATCTA
AGAGATTAGACGTTCCCTTCGGGGACATGGATACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGG
GTTAAGTCCCGCAACGAGCGCAACCCTTATTATCAGTTGCCAGCATTAAGTTGGGCACTCTGGTGAGACTGCCGGTG
ACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGA
TGGTACAACGAGTTGCGAACTCGCGAGAGTAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGATTGTAGGCTGCAAC
TCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACAC
ACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGTCGGTGGGGTAACCTTTTAGGAACCAGCCGCCTAAGTGA
CAGA
PheS result of gene sequence determination:
GGCGACAATTACAAGACGTGCTACTACGCACGCAGACGTCTGCTGATCAGCCGCGGTCACTTGAAAATC
ACGATTTTTCTAAAGGACCGCTGAAGGTCTTGTCACCTGGCCGCGTTTATCGGCGTGATACGGATGATGCAACCCAT
TCCCATCAATTTCATCAAATTGAAGGGTTAGTCGTGGACAAGCATATTACGATGGCTGATTTGAAGGGCACCTTAAT
TCTGGTTGCCAAGACTTTGTTTGGCGATCAATTCGATGTTCGGCTACGGCCAAGCTTCTTTCCATTCACGGAACCAT
CCGTAGAAGCTGATGTAACTTGCTTTAATTGCAATGGCAAGGGCTGTGCAATCTGTAAGCAAACGGGTTGGATCGAA
GTACTGGGTGCCGGCATGGTTCACCCCCACGTGTTAGAAATGTCTGGCATTGATCCAGAAGAATATGGTGGTTTTGC
TTTCGGTCTAGGGA
The measurement of 2 lactobacillus plantarum EM-12 thalli growth performance of embodiment and product acid activity
(1) preparation of culture medium:
It takes a certain amount of cereal materials to be crushed to 1.0mm, adds water to mix in the ratio of material water weight ratio 1:3, then will mix
Feed liquid high-temperature sterilization is closed, then is cooled to 36 DEG C or so, obtains fermentation medium.Wherein, the sterilising conditions are 121 DEG C of sterilizings,
Time 20 minutes.The cereal materials include rice meal, soybean powder and corn flour, the rice flour, soybean powder and corn flour
Weight ratio is 1:1:1.
(2) acquisition of fermenting microbe:
Take the glycerol tube preservation lactobacillus strain, be seeded to MRS culture medium, 36 DEG C stationary culture 20 hours, obtain
Lactobacillus strain.Wherein, the MRS fluid nutrient medium composition is as follows: 10g peptone, 5g beef extract, 4g yeast extract, the Portugal 20g
Grape sugar, 2g dipotassium hydrogen phosphate, 5g sodium acetate, 2g trisodium citrate, 1mL Tween 80,0.2g magnesium sulfate, 0.05g manganese sulfate are added
1000mL distilled water adjusts pH to 6.5,121 DEG C of sterilizing 20min.
(3) by lactobacillus strain obtained by volume 2% inoculum concentration be inoculated in the fermentation medium of step (1)
In, 36 DEG C of standing Anaerobic culturels incubation time 20 hours, obtain fermentation liquid.After taking fermentation medium to dilute 10 times, in 600nm
Lower measurement absorbance value, absorbance value are 0.76 ± 0.02.Fermentation liquid centrifugation measurement pH and lactic acid content are taken, wherein pH value is
3.80 ± 0.05,7.5 ± 0.5g/L of lactic acid content.
The measurement of 2 lactobacillus plantarum type strain product acid activity of comparative example
China Committee for Culture Collection of Microorganisms's General Microbiological Culture collection lactobacillus plantarum type strain is taken,
Its product acid activity is measured according to (1), (2) and (3) the step of embodiment 2.
(1) with (1) the step of embodiment 2.
(2) with (2) the step of embodiment 2.
(3) after taking seed culture medium to dilute 10 times, absorbance value is measured at 600nm, absorbance value is 0.62 ± 0.04；
Fermentation liquid centrifugation measurement pH and lactic acid content are taken, wherein pH value is 4.10 ± 0.05,6.0 ± 0.5g/L of lactic acid content.
As it can be seen that lactobacillus plantarum EM-12 product acid activity with higher provided by the invention, can be used as acidulant addition
In feed.
The guaranteeing the quality of 3 lactobacillus plantarum EM-12 of embodiment, antipollution experiment
(1) non-fermented raw mixture is prepared: by weight percentage, by amino acid 2%, vitamin 0.5%, micro member
Element 0.5%, salt 1.0%, mountain flour 3.0%, calcium dihydrogen phosphate 3.0%, white granulated sugar 30% and the mixing of super steam fish meal 60%,
And being crushed to granularity is 1.0mm, and non-fermented raw mixture is made.
(2) fermentation liquid is prepared according to the step of embodiment 2.
(3) fermentation liquid and non-fermented raw mixture are mixed according to weight percent for 70%, 30%, is sent out
Ferment solution.
(4) it is packed using plastics Soft Roll, is sterilized using the general pasteurization of food service industry, sterilization time
Control was at 60 minutes.
(5) it guarantees the quality, antipollution experiment
1. the fermentation liquid of acquisition is poured into triangular flask, juxtaposition opening is placed in 30 DEG C of incubator；2. solution is fallen
Enter in aluminium foil bag, and heat sealing, is placed in 30 DEG C of incubators.Wherein, 1. in feed liquid keep 72 hours, free from extraneous odour, pH without
Variation.2. without swollen bag, free from extraneous odour, pH are unchanged for aluminium foil bag 72 hours in.
3 lactobacillus plantarum type strain of comparative example produce acid guarantee the quality, antipollution experiment
China Committee for Culture Collection of Microorganisms's General Microbiological Culture collection lactobacillus plantarum type strain is taken,
It is guaranteed the quality according to (1), (2), (3), (4) and (5) the step of embodiment 3, antipollution experiment.
(1)-(4) are the same as (1)-(4) the step of embodiment 3.
(5) it guarantees the quality, antipollution experiment
1. the fermentation liquid of acquisition is poured into triangular flask, juxtaposition opening is placed in 30 DEG C of incubator；2. solution is fallen
Enter in aluminium foil bag, and heat sealing, is placed in 30 DEG C of incubators.Wherein, 1. in feed liquid keep 72 hours, smell acid is smelly, pH
It is remarkably decreased.2. 72 hours obvious swollen bags of aluminium foil bag in, smell acid is smelly, pH is remarkably decreased.
As it can be seen that lactobacillus plantarum EM-12 provided by the invention have preferably guarantee the quality, contamination resistance.
It should be noted that in another embodiment, the preparation of the non-fermented raw mixture the following steps are included:
By weight percentage, by amino acid 3%, vitamin 0.4%, microelement 0.6%, salt 1.5%, mountain flour 2.3%, phosphoric acid
Calcium dihydrogen 3.6%, white granulated sugar 38.6% and the mixing of super steam fish meal 50%, and being crushed to granularity is 1.2mm, is made non-
Fermentation raw material mixture.
The feeding effect of 4 fermented feed of embodiment
Fermented feed prepared by the above method is used for the feeding experiment of weanling pig, randomly chooses 100 health, weight
Uniform 28 age in days weanling pig, is divided into control group and test group, every group of 5 repetitions, each repetition 10, and experimental period 14 days.
Control group fed powder, feeding in 4 days is fermented liquid sucking pig material before test group, is gradually transited within the 5-7 days identical with control group
Powder, the powder identical with control group of feeding in the 8-14 days.Count the weight and diarrhea data such as following table of piglet.
As seen from the above table, piglet mostly weight gain 9.0% (1.29kg) of the piglet compared with feeding powder of liquid material, diarrhea rate are fed
Reduce by 70.6%.
The present invention can be obviously improved the immunity of livestock and poultry, improve intestine microenvironment, reduce diarrhea rate.
The fermentation situation of the collaboration yeast of embodiment 5
Fermenting microbe spreads cultivation: saccharomycete and lactobacillus plantarum EM-12 is selected, respectively through primary inclined plane culture, two-stage liquid
Body culture spreads cultivation, and steps are as follows:
(1) primary inclined plane culture
By lactobacillus switching in MRS medium slant, cultivated 24 hours in 36 DEG C；Saccharomyces cerevisiae switching is trained in YPD
It supports on base inclined-plane, is cultivated 20 hours in 30 DEG C；
(2) secondary liquid culture
The fresh lactobacillus inclined plane inoculating for taking step (1) to obtain is cultivated in fluid nutrient medium, is sealed with sterile sealed membrane,
Be placed in constant incubator, 36 DEG C of stationary culture 20h obtain level-one lactobacillus seed liquor, take primary seed solution in 5% ratio
It is seeded in secondary seed medium, the same first order seed of condition of culture obtains the seed liquor of lactobacillus；
The fresh saccharomyces cerevisiae inclined-plane for taking step (1) to obtain is forwarded in fluid nutrient medium, 30 DEG C shaking table culture 18 hours,
Level-one saccharomyces cerevisiae seed liquor is obtained, primary seed solution is taken to be inoculated in secondary seed medium by 5%, the same level-one of condition of culture
Seed；Obtain the seed liquor of saccharomyces cerevisiae.
(3) preparation of culture medium:
It takes a certain amount of cereal materials to be crushed to 1.0mm, adds water to mix in the ratio of material water weight ratio 1:4, by mixture
Liquid high-temperature sterilization, and 36 DEG C or so are cooled to, obtain fermentation medium.Wherein, the sterilising conditions are 121 DEG C of sterilizings, time
20 minutes.The cereal materials include rice meal, soybean powder and corn flour, the weight of the rice flour, soybean powder and wheat flour
Amount is than being 3:1:1.
(4) seed liquor, the seed liquor of lactobacillus strain for taking saccharomyces cerevisiae respectively, are connect by the inoculum concentration of total inoculum concentration 4%
Enter same fermentation medium.34 DEG C of cultivation temperature, shaking speed 200rpm.Wherein, saccharomycete and lactobacillus are inoculated with volume ratio 1:
1.Fermentation 20 hours, yeast viable count are 1.1 × 108Cfu/ml, living preparation of lactobacillus number 2.2 × 109cfu/ml。
In the present embodiment, the saccharomycete is saccharomyces cerevisiae (Saccharomyces cerevisiae) Sa-10, has been protected
It is hidden in China Committee for Culture Collection of Microorganisms's common micro-organisms center, CGMCC No.6120.
Comparative example 5
Lactobacillus plantarum EM-12 is replaced with into China Committee for Culture Collection of Microorganisms's General Microbiological Culture preservation
Center lactobacillus plantarum type strain, other steps are the same as embodiment 5.Fermentation 20 hours, yeast viable count are 0.8 × 108Cfu/ml,
Living preparation of lactobacillus number 1.8 × 109cfu/ml。
As it can be seen that the bacterial strain has more superior growth performance when the probiotics for cooperateing with yeast class carries out fermented and cultured, because
This is more suitable for the cooperative fermentation with yeast class strain, and having when preparing multiple bacteria compound fermentation feed more has research and development, makes
Use potentiality.
Strain provided by the invention growth, produce acid, in terms of show good performance, fermented product has
Good feeding effect.It, which is metabolized lactic acid, can inhibit enteric pathogenic bacteria growth, so as to be obviously improved the immunity of livestock and poultry, change
Kind intestine microenvironment, reduces disease incidence, and bring certain promotion to digestibility, the output of milk etc..
It should be understood by those ordinary skilled in the art that: the discussion of any of the above embodiment is exemplary only, not
It is intended to imply that the scope of the present disclosure (including claim) is limited to these examples；Under thinking of the invention, above embodiments
Or can also be combined between the technical characteristic in different embodiments, step can be realized with random order, and be existed such as
Many other variations of the upper different aspect of the invention, for simplicity, they are not provided in details.Therefore, it is all
Within the spirit and principles in the present invention, any omission, modification, equivalent replacement, improvement for being made etc. be should be included in of the invention
Within protection scope.
Sequence table
16S rDNA sequencing results:
GCGTGCTAATACATGCAGTCGAACGAACTCTGGTATTGATTGGTGCTTGCATCATGATTTACATTTGAG
TGAGTGGCGAACTGGTGAGTAACACGTGGGAAACCTGCCCAGAAGCGGGGGATAACACCTGGAAACAGATGCTAATA
CCGCATAACAACTTGGACCGCATGGTCCGAGTTTGAAAGATGGCTTCGGCTATCACTTTTGGATGGTCCCGCGGCGT
ATTAGCTAGATGGTGGGGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGAGAGGGTAATCGGCCACATTGG
GACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCA
ACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAACATATCTGAGAGTAACTGTTC
AGGTATTGACGGTATTTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCG
TTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCTTCGGCTCAACCGAA
GAAGTGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGA
TATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGTATGGGTAGCAA
ACAGGATTAGATACCCTGGTAGTCCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCT
GCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGGCCGCAAGGCTGAAACTCAAAGGAATTGACGGGGGCCCGC
ACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATACTATGCAAATCTA
AGAGATTAGACGTTCCCTTCGGGGACATGGATACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGG
GTTAAGTCCCGCAACGAGCGCAACCCTTATTATCAGTTGCCAGCATTAAGTTGGGCACTCTGGTGAGACTGCCGGTG
ACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGA
TGGTACAACGAGTTGCGAACTCGCGAGAGTAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGATTGTAGGCTGCAAC
TCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACAC
ACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGTCGGTGGGGTAACCTTTTAGGAACCAGCCGCCTAAGTGA
CAGA
PheS result of gene sequence determination:
GGCGACAATTACAAGACGTGCTACTACGCACGCAGACGTCTGCTGATCAGCCGCGGTCACTTGAAAATC
ACGATTTTTCTAAAGGACCGCTGAAGGTCTTGTCACCTGGCCGCGTTTATCGGCGTGATACGGATGATGCAACCCAT
TCCCATCAATTTCATCAAATTGAAGGGTTAGTCGTGGACAAGCATATTACGATGGCTGATTTGAAGGGCACCTTAAT
TCTGGTTGCCAAGACTTTGTTTGGCGATCAATTCGATGTTCGGCTACGGCCAAGCTTCTTTCCATTCACGGAACCAT
CCGTAGAAGCTGATGTAACTTGCTTTAATTGCAATGGCAAGGGCTGTGCAATCTGTAAGCAAACGGGTTGGATCGAA
GTACTGGGTGCCGGCATGGTTCACCCCCACGTGTTAGAAATGTCTGGCATTGATCCAGAAGAATATGGTGGTTTTGC
TTTCGGTCTAGGGA
Claims (8)
1. a lactobacillus plantarum (Lactobacillus plantarum), which is characterized in that it is micro- that the bacterial strain is preserved in China
Biological inoculum preservation administration committee common micro-organisms center, preservation place are in Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Institute of microbiology of the academy of sciences of state, the number that preservation is registered on the books are CGMCC No.11721, and the deposit date is in November, 2015
23 days.
2. a kind of lactobacillus plantarum as described in claim 1 (Lactobacillus plantarum) is in fermented feed
Using.
3. application according to claim 2 characterized by comprising
Into cereal materials plus water mixes, and fermentation medium is made, by the lactobacillus plantarum (Lactobacillus
Plantarum it) is inoculated in the fermentation medium and ferments, fermentation liquid is made；
Non-fermented raw mixture is mixed with fermentation liquid, fermented feed is made；Wherein, the non-fermented raw mixture includes
Amino acid, vitamin, microelement, salt, mountain flour, calcium dihydrogen phosphate, white granulated sugar and super steam fish meal.
4. application according to claim 3, which is characterized in that by the lactobacillus plantarum (Lactobacillus
Plantarum it) is all inoculated in the fermentation medium and is assisted with saccharomyces cerevisiae (Saccharomyces cerevisiae)
With fermentation.
5. application according to claim 4, which is characterized in that the saccharomycete is saccharomyces cerevisiae (Saccharomyces
Cerevisiae) Sa-10 has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, CGMCC
No.6120。
6. application according to claim 3, which is characterized in that the cereal materials are popcorn, dehulled soybean meal or hair
Ferment dregs of beans, sweet potato powder or soybean powder, rice flour or small rice and flour, wheat flour, the popcorn, dehulled soybean meal or fermentation beans
The weight percentage of the dregs of rice, sweet potato powder or soybean powder, rice flour or small rice and flour, wheat flour and water is followed successively by 1~20%, 1~
50%, 1~20%, 1~20%, 1~20% and 50~90%.
7. application according to claim 3, which is characterized in that the cereal materials be rice flour, soybean powder and corn flour,
The weight percentage of the rice flour, soybean powder and corn flour is followed successively by 1~30%, 1~30% and 40~80%.
8. application according to claim 3 or 4, which is characterized in that the lactobacillus plantarum (Lactobacillus
Plantarum inoculum concentration) is 1%~3%.
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| CN106578450A (en) * | 2016-11-25 | 2017-04-26 | 南宁学院 | Micro-ecologic preparation for improving immunity of calves, and preparation method and application thereof |
| CN106937710A (en) * | 2017-04-07 | 2017-07-11 | 青岛农业大学 | It is adapted to the fermented feed and application method of mink furring period |
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| CN109022313B (en) * | 2018-08-02 | 2021-08-24 | 中国农业科学院兰州兽医研究所 | Lactobacillus plantarum |
| CN110283749A (en) * | 2019-07-02 | 2019-09-27 | 北京科拓恒通生物技术股份有限公司 | One plant of lactobacillus plantarum P-01 separated and its application |
| CN111004752B (en) * | 2019-12-30 | 2021-11-05 | 江南大学 | Ethanol-resistant lactobacillus plantarum and application thereof in fermented food |
| CN111685235B (en) * | 2020-06-24 | 2023-08-22 | 河南省科学院生物研究所有限责任公司 | Preparation method of lactobacillus plantarum fermented liquid complete feed for piglets |
| CN111593010B (en) * | 2020-06-30 | 2022-07-26 | 黄河三角洲京博化工研究院有限公司 | Lactobacillus plantarum and method for producing fermented feed by using same |
| CN115074261B (en) * | 2022-06-28 | 2023-07-14 | 四川轻化工大学 | Bradyyeast, fermentation medium thereof, ruminant feed and preparation method of ruminant feed |
| CN116083273B (en) * | 2022-09-05 | 2024-04-19 | 新希望六和股份有限公司 | Lactobacillus plantarum NHE-LpE and application thereof |
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Denomination of invention: A Plant Lactobacillus Strain and Its Application in Fermented Feed Granted publication date: 20181218 Pledgee: Beijing technology intellectual property financing Company limited by guarantee Pledgor: BEIJING ENHALOR INTERNATIONAL TECH CO.,LTD. Registration number: Y2024990000166 |