CN111100808B - 一株植物乳杆菌及其应用 - Google Patents
一株植物乳杆菌及其应用 Download PDFInfo
- Publication number
- CN111100808B CN111100808B CN201911211085.7A CN201911211085A CN111100808B CN 111100808 B CN111100808 B CN 111100808B CN 201911211085 A CN201911211085 A CN 201911211085A CN 111100808 B CN111100808 B CN 111100808B
- Authority
- CN
- China
- Prior art keywords
- lactobacillus plantarum
- strain
- feed
- bacteria
- piglets
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 240000006024 Lactobacillus plantarum Species 0.000 title claims abstract description 30
- 235000013965 Lactobacillus plantarum Nutrition 0.000 title claims abstract description 30
- 229940072205 lactobacillus plantarum Drugs 0.000 title claims abstract description 30
- 238000000034 method Methods 0.000 claims abstract description 8
- 238000002360 preparation method Methods 0.000 claims description 16
- 230000001580 bacterial effect Effects 0.000 claims description 12
- 238000004321 preservation Methods 0.000 claims description 11
- 238000011081 inoculation Methods 0.000 claims description 7
- 235000015097 nutrients Nutrition 0.000 claims description 6
- 240000002791 Brassica napus Species 0.000 claims description 5
- 235000004977 Brassica sinapistrum Nutrition 0.000 claims description 5
- 230000000813 microbial effect Effects 0.000 claims description 5
- 238000010563 solid-state fermentation Methods 0.000 claims description 5
- 239000003674 animal food additive Substances 0.000 claims description 4
- 239000003795 chemical substances by application Substances 0.000 claims description 3
- 239000002994 raw material Substances 0.000 claims description 3
- 239000000126 substance Substances 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 230000012010 growth Effects 0.000 abstract description 18
- 238000004519 manufacturing process Methods 0.000 abstract description 13
- 239000002253 acid Substances 0.000 abstract description 11
- 239000003242 anti bacterial agent Substances 0.000 abstract description 10
- 229940088710 antibiotic agent Drugs 0.000 abstract description 10
- 210000001035 gastrointestinal tract Anatomy 0.000 abstract description 10
- 230000004083 survival effect Effects 0.000 abstract description 6
- 206010012735 Diarrhoea Diseases 0.000 abstract description 3
- 231100000956 nontoxicity Toxicity 0.000 abstract description 3
- 230000002349 favourable effect Effects 0.000 abstract description 2
- 230000008569 process Effects 0.000 abstract description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 26
- 241000894006 Bacteria Species 0.000 description 22
- 235000014655 lactic acid Nutrition 0.000 description 13
- 239000004310 lactic acid Substances 0.000 description 13
- 239000007788 liquid Substances 0.000 description 11
- 239000001963 growth medium Substances 0.000 description 10
- 238000012258 culturing Methods 0.000 description 9
- 239000006228 supernatant Substances 0.000 description 9
- 239000003833 bile salt Substances 0.000 description 8
- 244000052616 bacterial pathogen Species 0.000 description 5
- 230000003115 biocidal effect Effects 0.000 description 5
- 230000000529 probiotic effect Effects 0.000 description 5
- 230000003213 activating effect Effects 0.000 description 4
- 230000029087 digestion Effects 0.000 description 4
- 210000004051 gastric juice Anatomy 0.000 description 4
- 230000000968 intestinal effect Effects 0.000 description 4
- 239000002504 physiological saline solution Substances 0.000 description 4
- 239000006041 probiotic Substances 0.000 description 4
- 235000018291 probiotics Nutrition 0.000 description 4
- 230000035945 sensitivity Effects 0.000 description 4
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- 241000607142 Salmonella Species 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 108020000946 Bacterial DNA Proteins 0.000 description 2
- ULGZDMOVFRHVEP-RWJQBGPGSA-N Erythromycin Chemical compound O([C@@H]1[C@@H](C)C(=O)O[C@@H]([C@@]([C@H](O)[C@@H](C)C(=O)[C@H](C)C[C@@](C)(O)[C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)(C)O)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 ULGZDMOVFRHVEP-RWJQBGPGSA-N 0.000 description 2
- 230000005526 G1 to G0 transition Effects 0.000 description 2
- 238000003794 Gram staining Methods 0.000 description 2
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 2
- 241000186660 Lactobacillus Species 0.000 description 2
- 241000282887 Suidae Species 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 230000003385 bacteriostatic effect Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- MYSWGUAQZAJSOK-UHFFFAOYSA-N ciprofloxacin Chemical compound C12=CC(N3CCNCC3)=C(F)C=C2C(=O)C(C(=O)O)=CN1C1CC1 MYSWGUAQZAJSOK-UHFFFAOYSA-N 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000002496 gastric effect Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 229940039696 lactobacillus Drugs 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 244000144972 livestock Species 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 244000144977 poultry Species 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 229960005322 streptomycin Drugs 0.000 description 2
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- GSDSWSVVBLHKDQ-UHFFFAOYSA-N 9-fluoro-3-methyl-10-(4-methylpiperazin-1-yl)-7-oxo-2,3-dihydro-7H-[1,4]oxazino[2,3,4-ij]quinoline-6-carboxylic acid Chemical compound FC1=CC(C(C(C(O)=O)=C2)=O)=C3N2C(C)COC3=C1N1CCN(C)CC1 GSDSWSVVBLHKDQ-UHFFFAOYSA-N 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 108091093088 Amplicon Proteins 0.000 description 1
- 208000031295 Animal disease Diseases 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 108010062877 Bacteriocins Proteins 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- 241001478240 Coccus Species 0.000 description 1
- 108010078777 Colistin Proteins 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 241000282414 Homo sapiens Species 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 235000019779 Rapeseed Meal Nutrition 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 229960003022 amoxicillin Drugs 0.000 description 1
- LSQZJLSUYDQPKJ-NJBDSQKTSA-N amoxicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=C(O)C=C1 LSQZJLSUYDQPKJ-NJBDSQKTSA-N 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000009635 antibiotic susceptibility testing Methods 0.000 description 1
- 235000014590 basal diet Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229940093761 bile salts Drugs 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- UDSAIICHUKSCKT-UHFFFAOYSA-N bromophenol blue Chemical compound C1=C(Br)C(O)=C(Br)C=C1C1(C=2C=C(Br)C(O)=C(Br)C=2)C2=CC=CC=C2S(=O)(=O)O1 UDSAIICHUKSCKT-UHFFFAOYSA-N 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 210000004913 chyme Anatomy 0.000 description 1
- 229960003405 ciprofloxacin Drugs 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 229960001127 colistin sulfate Drugs 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- NJCUSQKMYNTYOW-MWUYRYRWSA-N enramicina Chemical compound O.N1C(=O)NC(=O)C(C=2C=C(Cl)C(O)=C(Cl)C=2)NC(=O)C(CO)NC(=O)C(C=2C=CC(O)=CC=2)NC(=O)C(CC2N=C(N)NC2)NC(=O)C(CCCNC(N)=O)NC(=O)C(C(C)O)NC(=O)C(C=2C=CC(O)=CC=2)NC(=O)C(C=2C=CC(O)=CC=2)NC(=O)C(C(C)O)NC(=O)N(CCCCN)C(=O)C(C=2C=CC(O)=CC=2)NC(=O)C(NC(=O)C(CC(O)=O)NC(=O)/C=C/C=C/CCCCC(C)CC)C(C)OC(=O)C(C=2C=CC(O)=CC=2)NC(=O)C(C)NC(=O)C1CC1CNC(N)=N1 NJCUSQKMYNTYOW-MWUYRYRWSA-N 0.000 description 1
- 229950003984 enramycin Drugs 0.000 description 1
- 108700041171 enramycin Proteins 0.000 description 1
- 229960003276 erythromycin Drugs 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 244000005709 gut microbiome Species 0.000 description 1
- 229910000037 hydrogen sulfide Inorganic materials 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 1
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 1
- 230000007413 intestinal health Effects 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000012982 microporous membrane Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000006872 mrs medium Substances 0.000 description 1
- ZESIAEVDVPWEKB-ORCFLVBFSA-N n-[(2s)-4-amino-1-[[(2s,3r)-1-[[(2s)-4-amino-1-oxo-1-[[(3s,6s,9s,12s,15r,18s,21s)-6,9,18-tris(2-aminoethyl)-3-[(1r)-1-hydroxyethyl]-12,15-bis(2-methylpropyl)-2,5,8,11,14,17,20-heptaoxo-1,4,7,10,13,16,19-heptazacyclotricos-21-yl]amino]butan-2-yl]amino]-3-h Chemical compound OS(O)(=O)=O.OS(O)(=O)=O.CC(C)CCCCC(=O)N[C@@H](CCN)C(=O)N[C@H]([C@@H](C)O)CN[C@@H](CCN)C(=O)N[C@H]1CCNC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCN)NC(=O)[C@H](CCN)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](CC(C)C)NC(=O)[C@H](CCN)NC1=O.CCC(C)CCCCC(=O)N[C@@H](CCN)C(=O)N[C@H]([C@@H](C)O)CN[C@@H](CCN)C(=O)N[C@H]1CCNC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCN)NC(=O)[C@H](CCN)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](CC(C)C)NC(=O)[C@H](CCN)NC1=O ZESIAEVDVPWEKB-ORCFLVBFSA-N 0.000 description 1
- 229960001180 norfloxacin Drugs 0.000 description 1
- OGJPXUAPXNRGGI-UHFFFAOYSA-N norfloxacin Chemical compound C1=C2N(CC)C=C(C(O)=O)C(=O)C2=CC(F)=C1N1CCNCC1 OGJPXUAPXNRGGI-UHFFFAOYSA-N 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 229960001699 ofloxacin Drugs 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- LSQZJLSUYDQPKJ-UHFFFAOYSA-N p-Hydroxyampicillin Natural products O=C1N2C(C(O)=O)C(C)(C)SC2C1NC(=O)C(N)C1=CC=C(O)C=C1 LSQZJLSUYDQPKJ-UHFFFAOYSA-N 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 238000004080 punching Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000004456 rapeseed meal Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 150000003839 salts Chemical group 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
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
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
-
- 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
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- Animal Husbandry (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Physiology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- General Engineering & Computer Science (AREA)
- Birds (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Sustainable Development (AREA)
- Virology (AREA)
- General Health & Medical Sciences (AREA)
- Botany (AREA)
- Mycology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Fodder In General (AREA)
Abstract
本发明公开了一株植物乳杆菌及其应用。本发明提供菌株筛选自猪胃肠道，能够很好的耐受胃肠道的环境，有利于菌株在肠道中存活、增殖和定植；本发明提供的菌株特异性强，生长速度快、产酸高、抑菌、无毒害；使用本发明提供的菌株能够降低仔猪的腹泻率，提高其生长性能；在生产上使用本发明提供菌株能够减少饲喂过程中抗生素的使用量。
Description
技术领域
本发明属于微生物领域，涉及一株植物乳杆菌及其应用。
背景技术
我国是畜禽养殖大国，也是抗生素生产和使用大国。抗生素在防治动物疾病、提高养殖效益的同时，抗生素滥用问题也普遍存在。因此，寻找绿色环保型抗生素替代物，减轻抗生素对畜禽生产带来的负面影响成为我国畜牧业迫切需要解决的问题。益生菌由于其具有绿色、安全等特点，成为目前饲用抗生素的主要替代品。但在实际生产中，一些益生菌制剂由于其活性太低且对环境的耐受性较差等，并不能稳定的发挥其益生作用。植物乳杆菌是人和动物肠道中的正常菌群，已有大量研究表明植物乳杆菌能够抑制病原菌(如大肠杆菌、沙门氏菌)增殖，促进机体对营养物质的消化吸收，提高机体免疫力等。植物乳杆菌通过与病原菌对限制性营养物质的竞争，从而抑制病原菌的生长，调节肠道微生物区系的组成，改善肠道健康。同时，通过其生长代谢所产生的乳酸、细菌素等对肠道中其他微生物和宿主的左右，调节菌群之间的关系，使肠道有优势菌保持稳定的数量和状态，阻止病原菌的定植和入侵，是一种理想的微生物制剂。因此植物乳杆菌在畜牧生产中具有广泛的应用前景。
目前在猪生产中主要采用抗生素治疗仔猪腹泻、提高生长性能，而结果会导致猪肠道菌群的失调。而微生态制剂的使用能够抑制肠道病原菌的增殖，并提高有益菌的数量。植物乳杆菌是生产中常用的微生态制剂，但由于市场上大多数植物乳杆菌制剂对仔猪的益生效果不明显。因此，本发明的目的是提供一株具有益生性能，在仔猪生产上能够替代抗生素使用的植物乳杆菌制剂，
并提供该菌株在仔猪生产中替代抗生素使用的方法。
发明内容
本发明的目的是针对现有技术的上述不足，提供一株植物乳杆菌及其应用。。
本发明的目的可通过以下技术方案实现：
一株植物乳杆菌(lactobacillus plantarum)L47，保藏于中国典型培养物保藏中心，保藏日期为2019年11月22日，保藏编号为CCTCC NO:M2019967。
由植物乳杆菌(lactobacillus plantarum)L47制备的菌剂。
一种饲料添加剂，含有所述的L47的菌剂。
所述的植物乳杆菌(lactobacillus plantarum)L47在制备饲料添加剂中的应用。
所述的植物乳杆菌(lactobacillus plantarum)L47在发酵制备饲料中的应用。
一种猪饲料的制备方法，L47菌株经过活化后，按1×108CFU/g原料的接种量接种至料水比为1:1.1的菜籽粕中，在37℃条件下发酵48小时，使L47在不添加其他营养物质的情况下固态发酵48h得到所述的猪饲料。
有益效果：
本发明提供菌株筛选自猪胃肠道，能够很好的耐受胃肠道的环境，有利于菌株在肠道中存活、增殖和定植；本发明提供的菌株特异性强，生长速度快、产酸高、抑菌、无毒害；使用本发明提供的菌株能够降低仔猪的腹泻率，提高其生长性能；.在生产上使用本发明提供菌株能够减少饲喂过程中抗生素的使用量。
1.本发明提供的菌株特异性强，生长速度快、产酸高、抑菌、无毒害；
2.本发明提供菌株在MRS培养基上生长20h其活菌数能达到1.0×109cfu/mL；.
3.本发明提供菌株制剂适用于断奶仔猪，且在仔猪饲料中的添加量为2.0×108cfu/kg。
附图说明
图1筛选出的4株乳酸菌的生长曲线
图2菌株在人工模拟胃肠液下的存活率
图3本发明技术流程图
生物材料保藏信息
L47,分类命名为植物乳杆菌L47 Lactobacillus plantarum L47,保藏于中国典型培养物保藏中心，保藏地址为中国武汉.武汉大学,保藏日期为2019年11月22日，保藏编号为CCTCC NO:M2019967。
具体实施方式
实施例1植物乳杆菌的筛选和鉴定
1.1样品来源、分离和纯化
本发明所提供的植物乳杆菌分离自不同生理阶段猪(仔猪、生长育肥猪和母猪)肠道粘膜、食糜及粪便。将采集样品进行梯度稀释(1×10-4～1×10-6)后，在MRS固体培养基上涂布，37℃培养24-36h，挑取单菌落，经2-3次纯化后，将培养液置于40％甘油，于-20℃保存。
1.2菌株形态观察
取少量保存菌液，充分活化。将活化菌液涂于事先滴有少量无菌生理盐水的载玻片上，将玻片涂匀，然后待玻片干燥、固定后，对细菌进行革兰氏染色及镜检，最后观察菌体形态特征。共分离得到猪源乳酸菌155株，其中杆菌105株，球菌50株。革兰氏染色呈阳性(镜检呈蓝色)，单个、成对或链状排列。
1.3菌株产酸能力测定
将保存的菌种接种至10mL标准MRS肉汤中，37℃厌氧培养24h，充分活化。将活化的100μL菌液接种至改良MRS培养基1(含0.01g/L溴酚蓝指示剂的MRS培养基)30℃厌氧培养，记录培养物的颜色(由紫色变为黄色)的时间，在90h后测定菌液pH值。选择产酸速度快(变色时间短)、终pH值较低的乳酸菌，测定其乳酸含量，取100μL活化后的菌液接种至改良MRS培养基2(含2％CaCO3的MRS培养基)中，30℃条件下培养48h后取样，测定乳酸含量。每个菌株3个重复。
1.4菌株鉴定
1.4.1菌株常规生化鉴定
对候选菌株进行接触酶、硫化氢、明胶、吲哚、硝酸盐还原试验，进行10℃、45℃、60℃生长试验，6.5％NaCl生长试验，pH 9.6生长试验。
1.4.2菌株16S rRNA鉴定
将保存菌液充分活化后，95℃对菌体进行裂解，例行获取细菌DNA，以细菌DNA为模版，使用细菌全序列通用引物8f(5’AGA GTT TGA TCC TGG CTC AG3’)和1510r(5’GGC TACCTT GTT ACG A3’)对候选菌株DNA进行PCR扩展，应用琼脂糖(1.2％)凝胶电泳对扩增产物进行检测，扩增子约为1400bp，之后送Invitrogen公司进行测序，将所得序列提交GenBank进行Blast比对分析。初步分离出4株具有良好产酸性能的乳酸菌。
1.5菌株生长曲线的绘制
按照前述方法活化菌株，按1％的接种量将菌液接种到MRS液体培养基中。37℃进行培养，从培养0小时开始，每2小时取样一次，对菌液的吸光值(OD600)进行测定，直至20小时结束。绘制出生长曲线(以培养时间为横坐标，吸光值为纵坐标)。生长曲线如图1所示。4株候选菌株在接种后4h快速进入对数生长期，其中L47、L63、L79在接种12h后进入生长稳定期，L47在14h后进入生长稳定期，这与前人结果基本一致。说明该4株乳酸菌可在营养丰富的环境下快速生长，满足优良益生菌的特性。
实施例2.菌株益生性能检测
2.1菌株耐酸、耐胆盐性能
耐酸性能：将候选菌株接种于MRS培养基中，37℃条件下培养24h，活化后，每株菌株分别取两等份菌液4mL，离心(10000r/min，5min)，弃上清，加入4mL已灭菌的pH 2.5和6.2(空白对照)的生理盐水，37℃，2h后，取400μL接种到10mL的MRS液体培养基中，培养20h后，测定吸光值(OD600)。
耐胆盐性能：与耐酸性能测试相似，活化菌株，离心，弃上清，加入4mL已灭菌0.5％胆盐或0％胆盐溶液(空白对照)，37℃保温2h后取样，接种，20h后测定OD600(操作与耐酸性能试验完全一样)。
菌株L47在pH 6.2(对照组)和pH 2.5条件下OD600值差异不显著(P>0.05)，说明该株乳酸菌能够耐受较低的pH。4株乳酸菌中，L47在0.5％胆盐组的OD600值与对照组差异不显著(P>0.05)，说明该乳酸菌能够耐受0.5％胆盐。
表1筛选出的4株乳酸菌对酸和胆盐的耐受性
注在同行不同字母表示差异显著(P＜0.05)，相同字母表示差异不显著(P＞0.05)
2.2菌株上清液抑菌性能
将候选菌株制成3×108CFU/mL的悬液于MRS液体培养基中37℃培养活化2代，将活化好的菌株离心(10000g，4℃，10min)，上清液经滤膜(0.22μm)过滤，收集后分为等量的2份，一份不调pH，另一份将pH调至6.5，检测有机酸是否对指示菌产生抑制作用。指示菌大肠杆菌K88接种于LB培养基中37℃培养24h，离心(10000g，4℃，10min)，弃上清，菌体悬于灭菌生理盐水，离心洗涤1次，制成3×108CFU/mL菌悬液，取200μL接种到琼脂培养基中，静置2h，待琼脂板凝固后，牛津杯(直径8mm)在上面打孔。分别取上述不调pH和pH 6.5的上清液100μL加入孔中，室温静置3-5h，37℃培养48h，测量抑菌圈直径，每组3个重复。沙门氏菌操作方法同上。L47上清液对指示菌的抑制作用最强。
表2筛选出4株乳酸菌对E.coli K88和沙门氏菌的抑菌性能
注：NE表示未抑制
2.3抗生素敏感性试验
采用纸片扩散法测定菌株的抗生素敏感性。稀释后的菌液(100μL；6.0log CFU/mL)均匀的涂布于直径为90mm的MRS琼脂培养基表面，使用无菌镊子将含有抗生素的药敏纸片放至平板表面，包括氧氟沙星、阿莫西林、环丙沙星、诺氟沙星、红霉素、链霉素和氨苄西林。37℃培养24h后对抑菌圈直径进行测定。L47除对链霉素耐药外，对其余几种抗生素表现出敏感或中介敏感。
表3菌株的药敏试验结果
注：S：敏感，I：中介敏感，R：耐药。
Note：S:Susceptible,I:Intermediate,R:Resistant.
2.4模拟胃肠道消化试验
人工胃液：在pH为2.5的无菌生理盐水中，分别加入3.0g/L胃蛋白酶，充分溶解后，用无菌微孔滤膜(0.22μm)过滤除菌，得到人工胃液，备用。人工肠液：在pH为8.0的无菌生理盐水中，加入1.0g/L胰蛋白酶和1.8g/L胆盐，充分溶解后，过滤除菌备用。将活化后的菌液按10％(v/v)的接种量接入上述人工胃液中，于37℃培养4h；接入人工肠液中培养20h。采用涂布平板法对菌液进行活菌计数，并计算存活率。L47在pH 2.5的人工胃液消化4h，其存活率达到了99.11％，且在模拟肠液消化后存活率达到了60％。将L47送交CCTCC保藏，保藏编号为CCTCC M 2019967。
实施例3.植物乳杆菌制剂的应用
选取144头28日龄断奶仔猪，随机分为3组，对照组饲喂基础日粮，抗生素组在日粮中添加200mg/kg硫酸粘杆菌素和200mg/kg恩拉霉素，植物乳杆菌制剂组在日粮中添加2.0×108cfu/kg的活菌制剂。每组6个重复，每个重复8头仔猪。试验于32日龄开始，59日龄结束，共25天。其生长性能对比结果如表4所示。
表4植物乳杆菌对断奶仔猪生长性能的影响
4.植物乳杆菌发酵饲料
将植物乳杆菌L47作为接种物，对菜籽粕进行固态发酵，产生的乳酸等代谢产物能够降低菜籽粕的pH，抑制杂菌的生长。
菌株经过活化后，按1×108CFU/g原料的接种量接种至料水比为1:1.1的菜籽粕中，在37℃条件下发酵48小时，使乳酸杆菌在不添加其他营养物质的情况下固态发酵48h。结束后测定饲料的pH、乳酸和常规营养成分，结果见表5。
表5植物乳杆菌L47固态发酵菜籽粕
Claims (6)
1.一株植物乳杆菌（lactobacillus plantarum）L47，保藏于中国典型培养物保藏中心，保藏日期为2019年11月22日，保藏编号为CCTCC NO: M2019967。
2.由权利要求1所述的植物乳杆菌（lactobacillus plantarum）L47制备的菌剂。
3.一种饲料添加剂，其特征在于含有权利要求2所述的菌剂。
4.权利要求1所述的植物乳杆菌（lactobacillus plantarum）L47在制备饲料添加剂中的应用。
5.权利要求1所述的植物乳杆菌（lactobacillus plantarum）L47在发酵制备饲料中的应用。
6.一种猪饲料的制备方法，其特征在于将权利要求1所述的植物乳杆菌（lactobacillus plantarum）L47菌株经过活化后，按1×108 CFU/g 原料的接种量接种至料水比为1:1.1的菜籽粕中，在37℃条件下发酵48小时，使L47在不添加其他营养物质的情况下固态发酵48 h得到所述的猪饲料。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911211085.7A CN111100808B (zh) | 2019-12-02 | 2019-12-02 | 一株植物乳杆菌及其应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911211085.7A CN111100808B (zh) | 2019-12-02 | 2019-12-02 | 一株植物乳杆菌及其应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111100808A CN111100808A (zh) | 2020-05-05 |
| CN111100808B true CN111100808B (zh) | 2021-02-12 |
Family
ID=70421355
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911211085.7A Active CN111100808B (zh) | 2019-12-02 | 2019-12-02 | 一株植物乳杆菌及其应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111100808B (zh) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117603890A (zh) * | 2024-01-24 | 2024-02-27 | 山东大益生物科技集团有限公司 | 一种植物乳杆菌lp23菌株及其应用 |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105132322B (zh) * | 2015-09-02 | 2016-08-31 | 广州格拉姆生物科技有限公司 | 一株植物乳杆菌及其应用 |
| CN106906154B (zh) * | 2015-12-22 | 2021-07-27 | 北京大北农科技集团股份有限公司 | 一株植物乳杆菌菌株及其饲料添加剂与饲料 |
-
2019
- 2019-12-02 CN CN201911211085.7A patent/CN111100808B/zh active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN111100808A (zh) | 2020-05-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106282072B (zh) | 一种复合乳酸菌微生态制剂及其制备方法与应用 | |
| CN108251335B (zh) | 一种具有乳酸活性的粪肠球菌hkf7及其筛选培养方法和应用 | |
| CN115094012B (zh) | 凝结芽孢杆菌bc-hyc株菌剂的制备方法及应用 | |
| CN106497808A (zh) | 一株分离自猪肠道的可用于生产发酵饲料的唾液乳杆菌 | |
| CN116179440B (zh) | 一株鸡源贝莱斯芽孢杆菌及其应用 | |
| CN109136126B (zh) | 一株用于预防断奶仔猪腹泻的戊糖片球菌 | |
| CN114806978B (zh) | 约氏乳杆菌sxdt-23及其应用 | |
| CN111534447A (zh) | 约氏乳杆菌及其应用 | |
| CN106906166A (zh) | 一种猪源唾液乳杆菌活菌制剂及应用 | |
| CN110028560B (zh) | 一种凝结芽孢杆菌产的细菌素及其应用 | |
| CN114085789B (zh) | 戊糖片球菌ma.wtpqj01及其应用 | |
| CN108660097A (zh) | 一株鱼源屎肠球菌r8的筛选及应用 | |
| CN114836358A (zh) | 罗伊氏乳杆菌sxdt-32及其应用 | |
| CN114107137A (zh) | 一种丁酸梭菌及其应用和产品 | |
| CN111100808B (zh) | 一株植物乳杆菌及其应用 | |
| CN117448213A (zh) | 一种抑制产气荚膜梭菌的植物乳杆菌及其后生元和应用 | |
| CN116478873B (zh) | 一种民猪肠道中可抑制多种有害菌生长的贝莱斯芽孢杆菌 | |
| CN112226389A (zh) | 一种三黄肉雏鸡肠道益生菌群的定植培养方法及其应用 | |
| CN117327626A (zh) | 一种植物乳杆菌ts1及其培养方法与应用 | |
| CN114806944B (zh) | 植物乳杆菌lp11、其发酵液及制备方法和应用 | |
| CN114410514B (zh) | 斯密氏芽孢杆菌及其用途 | |
| CN114231464B (zh) | 凝结芽孢杆菌及其用途 | |
| CN116064324A (zh) | 一株鼠李糖乳杆菌及其培养方法和在防治腹泻和肠炎方面的应用 | |
| CN115572689A (zh) | 一株戊糖片球菌m6及其应用 | |
| CN115651860A (zh) | 凝结芽孢杆菌bc-hyc株及其应用 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |