CN104894031B - Leuconostoc mesenteroides and application thereof in low-temperature silage - Google Patents
Leuconostoc mesenteroides and application thereof in low-temperature silage Download PDFInfo
- Publication number
- CN104894031B CN104894031B CN201510340177.0A CN201510340177A CN104894031B CN 104894031 B CN104894031 B CN 104894031B CN 201510340177 A CN201510340177 A CN 201510340177A CN 104894031 B CN104894031 B CN 104894031B
- Authority
- CN
- China
- Prior art keywords
- leuconostoc mesenteroides
- ensilage
- fermentation
- leuconostoc
- mesenteroides
- 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.)
- Expired - Fee Related
Links
- 241000192130 Leuconostoc mesenteroides Species 0.000 title claims abstract description 142
- 239000004460 silage Substances 0.000 title claims abstract description 12
- 241000894006 Bacteria Species 0.000 claims abstract description 33
- 238000000034 method Methods 0.000 claims abstract description 9
- 238000004519 manufacturing process Methods 0.000 claims abstract description 3
- 230000001580 bacterial effect Effects 0.000 claims description 23
- 238000000855 fermentation Methods 0.000 claims description 16
- 230000004151 fermentation Effects 0.000 claims description 14
- 239000000654 additive Substances 0.000 claims description 9
- 230000000996 additive effect Effects 0.000 claims description 9
- 239000002068 microbial inoculum Substances 0.000 claims description 9
- 239000002994 raw material Substances 0.000 claims description 7
- 239000007787 solid Substances 0.000 claims description 6
- 241000191938 Micrococcus luteus Species 0.000 claims description 5
- 241000607142 Salmonella Species 0.000 claims description 4
- 230000001629 suppression Effects 0.000 claims description 4
- 239000003112 inhibitor Substances 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims 1
- 239000000835 fiber Substances 0.000 abstract description 24
- 235000019784 crude fat Nutrition 0.000 abstract description 10
- 235000015097 nutrients Nutrition 0.000 abstract description 10
- 235000019750 Crude protein Nutrition 0.000 abstract description 9
- 239000002253 acid Substances 0.000 abstract description 8
- 238000004321 preservation Methods 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 4
- 230000007774 longterm Effects 0.000 abstract description 3
- 150000003839 salts Chemical class 0.000 abstract description 3
- 230000008569 process Effects 0.000 abstract description 2
- 238000003860 storage Methods 0.000 abstract description 2
- 239000003513 alkali Substances 0.000 abstract 1
- 235000021095 non-nutrients Nutrition 0.000 abstract 1
- 239000000126 substance Substances 0.000 abstract 1
- 244000075850 Avena orientalis Species 0.000 description 26
- 235000007319 Avena orientalis Nutrition 0.000 description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 14
- 230000007935 neutral effect Effects 0.000 description 13
- 239000003599 detergent Substances 0.000 description 12
- 239000012530 fluid Substances 0.000 description 12
- 238000002474 experimental method Methods 0.000 description 11
- 239000011521 glass Substances 0.000 description 10
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 10
- 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 9
- 238000003556 assay Methods 0.000 description 9
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 8
- 239000001963 growth medium Substances 0.000 description 8
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid group Chemical class S(O)(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 8
- -1 D- turanoses Natural products 0.000 description 7
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 7
- 239000008103 glucose Substances 0.000 description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 6
- 229920001817 Agar Polymers 0.000 description 6
- 239000001888 Peptone Substances 0.000 description 6
- 108010080698 Peptones Proteins 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 239000008272 agar Substances 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 235000019319 peptone Nutrition 0.000 description 6
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 5
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 5
- 241000196324 Embryophyta Species 0.000 description 5
- 241000235342 Saccharomycetes Species 0.000 description 5
- 235000015278 beef Nutrition 0.000 description 5
- 229940041514 candida albicans extract Drugs 0.000 description 5
- 229910052799 carbon Inorganic materials 0.000 description 5
- 230000008859 change Effects 0.000 description 5
- 238000001035 drying Methods 0.000 description 5
- 239000000284 extract Substances 0.000 description 5
- 238000010438 heat treatment Methods 0.000 description 5
- 239000004310 lactic acid Substances 0.000 description 5
- 235000014655 lactic acid Nutrition 0.000 description 5
- 239000012138 yeast extract Substances 0.000 description 5
- 108020004465 16S ribosomal RNA Proteins 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 241000193403 Clostridium Species 0.000 description 4
- 241000588724 Escherichia coli Species 0.000 description 4
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 4
- 241000192132 Leuconostoc Species 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 4
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 4
- 235000019341 magnesium sulphate Nutrition 0.000 description 4
- 229940099596 manganese sulfate Drugs 0.000 description 4
- 239000011702 manganese sulphate Substances 0.000 description 4
- 235000007079 manganese sulphate Nutrition 0.000 description 4
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 4
- 235000013336 milk Nutrition 0.000 description 4
- 239000008267 milk Substances 0.000 description 4
- 210000004080 milk Anatomy 0.000 description 4
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 4
- 229920000053 polysorbate 80 Polymers 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- BDKLKNJTMLIAFE-UHFFFAOYSA-N 2-(3-fluorophenyl)-1,3-oxazole-4-carbaldehyde Chemical compound FC1=CC=CC(C=2OC=C(C=O)N=2)=C1 BDKLKNJTMLIAFE-UHFFFAOYSA-N 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 230000004913 activation Effects 0.000 description 3
- 239000001913 cellulose Substances 0.000 description 3
- 229920002678 cellulose Polymers 0.000 description 3
- 235000019504 cigarettes Nutrition 0.000 description 3
- 239000000470 constituent Substances 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 229940050410 gluconate Drugs 0.000 description 3
- 229910052573 porcelain Inorganic materials 0.000 description 3
- 235000017281 sodium acetate Nutrition 0.000 description 3
- 229940087562 sodium acetate trihydrate Drugs 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 3
- KBPLFHHGFOOTCA-UHFFFAOYSA-N 1-Octanol Chemical compound CCCCCCCCO KBPLFHHGFOOTCA-UHFFFAOYSA-N 0.000 description 2
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 2
- SRBFZHDQGSBBOR-SOOFDHNKSA-N D-ribopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@@H]1O SRBFZHDQGSBBOR-SOOFDHNKSA-N 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 235000014676 Phragmites communis Nutrition 0.000 description 2
- 238000010306 acid treatment Methods 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 238000004380 ashing Methods 0.000 description 2
- 230000003385 bacteriostatic effect Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 239000013530 defoamer Substances 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 239000003517 fume Substances 0.000 description 2
- 230000015784 hyperosmotic salinity response Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 229920005610 lignin Polymers 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 235000008935 nutritious Nutrition 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 230000000717 retained effect Effects 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 238000002798 spectrophotometry method Methods 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 238000000967 suction filtration Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000004448 titration Methods 0.000 description 2
- 238000009461 vacuum packaging Methods 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- DBOVHQOUSDWAPQ-UHFFFAOYSA-N (4aS)-6c-[O2-(3,5,3'-trihydroxy-biphenyl-2-carbonyl)-beta-D-glucopyranosyloxy]-5t-vinyl-(4ar)-4,4a,5,6-tetrahydro-3H-pyrano[3,4-c]pyran-1-one Natural products OC1C(O)C(CO)OC(OC2C(C3C(C(OCC3)=O)=CO2)C=C)C1OC(=O)C1=C(O)C=C(O)C=C1C1=CC=CC(O)=C1 DBOVHQOUSDWAPQ-UHFFFAOYSA-N 0.000 description 1
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- PLXMOAALOJOTIY-FPTXNFDTSA-N Aesculin Natural products OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](O)[C@H]1Oc2cc3C=CC(=O)Oc3cc2O PLXMOAALOJOTIY-FPTXNFDTSA-N 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- BZXINCMCFVKGKB-UHFFFAOYSA-N Amarogentin Natural products OCC1OC(OC2OC=C3C(CCOC3=O)C2C=C)C(OC(=O)c4cc(O)cc(O)c4c5cccc(O)c5)C(O)C1O BZXINCMCFVKGKB-UHFFFAOYSA-N 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 241000239290 Araneae Species 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 1
- 235000005979 Citrus limon Nutrition 0.000 description 1
- 244000131522 Citrus pyriformis Species 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- GUBGYTABKSRVRQ-CUHNMECISA-N D-Cellobiose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-CUHNMECISA-N 0.000 description 1
- RGHNJXZEOKUKBD-SQOUGZDYSA-M D-gluconate Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O RGHNJXZEOKUKBD-SQOUGZDYSA-M 0.000 description 1
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 1
- LKDRXBCSQODPBY-OEXCPVAWSA-N D-tagatose Chemical compound OCC1(O)OC[C@@H](O)[C@H](O)[C@@H]1O LKDRXBCSQODPBY-OEXCPVAWSA-N 0.000 description 1
- 206010013786 Dry skin Diseases 0.000 description 1
- 239000004278 EU approved seasoning Substances 0.000 description 1
- UNXHWFMMPAWVPI-UHFFFAOYSA-N Erythritol Natural products OCC(O)C(O)CO UNXHWFMMPAWVPI-UHFFFAOYSA-N 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 241000726221 Gemma Species 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 229920002527 Glycogen Polymers 0.000 description 1
- 229920002488 Hemicellulose Polymers 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- LKDRXBCSQODPBY-AMVSKUEXSA-N L-(-)-Sorbose Chemical compound OCC1(O)OC[C@H](O)[C@@H](O)[C@@H]1O LKDRXBCSQODPBY-AMVSKUEXSA-N 0.000 description 1
- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 description 1
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-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
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 1
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 1
- MBLBDJOUHNCFQT-LXGUWJNJSA-N N-acetylglucosamine Natural products CC(=O)N[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO MBLBDJOUHNCFQT-LXGUWJNJSA-N 0.000 description 1
- AYRXSINWFIIFAE-UHFFFAOYSA-N O6-alpha-D-Galactopyranosyl-D-galactose Natural products OCC1OC(OCC(O)C(O)C(O)C(O)C=O)C(O)C(O)C1O AYRXSINWFIIFAE-UHFFFAOYSA-N 0.000 description 1
- RCEAADKTGXTDOA-UHFFFAOYSA-N OS(O)(=O)=O.CCCCCCCCCCCC[Na] Chemical compound OS(O)(=O)=O.CCCCCCCCCCCC[Na] RCEAADKTGXTDOA-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 235000008331 Pinus X rigitaeda Nutrition 0.000 description 1
- 235000011613 Pinus brutia Nutrition 0.000 description 1
- 241000018646 Pinus brutia Species 0.000 description 1
- JVWLUVNSQYXYBE-UHFFFAOYSA-N Ribitol Natural products OCC(C)C(O)C(O)CO JVWLUVNSQYXYBE-UHFFFAOYSA-N 0.000 description 1
- PYMYPHUHKUWMLA-LMVFSUKVSA-N Ribose Natural products OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 description 1
- NGFMICBWJRZIBI-JZRPKSSGSA-N Salicin Natural products O([C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@H](CO)O1)c1c(CO)cccc1 NGFMICBWJRZIBI-JZRPKSSGSA-N 0.000 description 1
- DWAQJAXMDSEUJJ-UHFFFAOYSA-M Sodium bisulfite Chemical compound [Na+].OS([O-])=O DWAQJAXMDSEUJJ-UHFFFAOYSA-M 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- VLSOAXRVHARBEQ-UHFFFAOYSA-N [4-fluoro-2-(hydroxymethyl)phenyl]methanol Chemical compound OCC1=CC=C(F)C=C1CO VLSOAXRVHARBEQ-UHFFFAOYSA-N 0.000 description 1
- PQLVXDKIJBQVDF-UHFFFAOYSA-N acetic acid;hydrate Chemical class O.CC(O)=O PQLVXDKIJBQVDF-UHFFFAOYSA-N 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 241001148470 aerobic bacillus Species 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- HMFHBZSHGGEWLO-UHFFFAOYSA-N alpha-D-Furanose-Ribose Natural products OCC1OC(O)C(O)C1O HMFHBZSHGGEWLO-UHFFFAOYSA-N 0.000 description 1
- NGFMICBWJRZIBI-UHFFFAOYSA-N alpha-salicin Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=CC=C1CO NGFMICBWJRZIBI-UHFFFAOYSA-N 0.000 description 1
- DBOVHQOUSDWAPQ-WTONXPSSSA-N amarogentin Chemical compound O([C@H]1[C@H](O[C@H]2[C@@H]([C@H]3C(C(OCC3)=O)=CO2)C=C)O[C@@H]([C@H]([C@@H]1O)O)CO)C(=O)C1=C(O)C=C(O)C=C1C1=CC=CC(O)=C1 DBOVHQOUSDWAPQ-WTONXPSSSA-N 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- DLRVVLDZNNYCBX-ZZFZYMBESA-N beta-melibiose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](O)O1 DLRVVLDZNNYCBX-ZZFZYMBESA-N 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000000498 cooling water Substances 0.000 description 1
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 235000013325 dietary fiber Nutrition 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- UNXHWFMMPAWVPI-ZXZARUISSA-N erythritol Chemical compound OC[C@H](O)[C@H](O)CO UNXHWFMMPAWVPI-ZXZARUISSA-N 0.000 description 1
- XHCADAYNFIFUHF-TVKJYDDYSA-N esculin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC(C(=C1)O)=CC2=C1OC(=O)C=C2 XHCADAYNFIFUHF-TVKJYDDYSA-N 0.000 description 1
- 238000000895 extractive distillation Methods 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 235000021050 feed intake Nutrition 0.000 description 1
- 229960002413 ferric citrate Drugs 0.000 description 1
- 235000011194 food seasoning agent Nutrition 0.000 description 1
- 239000004459 forage Substances 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- FBPFZTCFMRRESA-GUCUJZIJSA-N galactitol Chemical compound OC[C@H](O)[C@@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-GUCUJZIJSA-N 0.000 description 1
- DLRVVLDZNNYCBX-CQUJWQHSSA-N gentiobiose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)C(O)O1 DLRVVLDZNNYCBX-CQUJWQHSSA-N 0.000 description 1
- 229930182478 glucoside Natural products 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 229940096919 glycogen Drugs 0.000 description 1
- BJRNKVDFDLYUGJ-RMPHRYRLSA-N hydroquinone O-beta-D-glucopyranoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=C(O)C=C1 BJRNKVDFDLYUGJ-RMPHRYRLSA-N 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 238000009413 insulation Methods 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- NPFOYSMITVOQOS-UHFFFAOYSA-K iron(III) citrate Chemical compound [Fe+3].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NPFOYSMITVOQOS-UHFFFAOYSA-K 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- ZBDGHWFPLXXWRD-JGWLITMVSA-N methyl beta-D-xylopyranoside Chemical compound CO[C@@H]1OC[C@@H](O)[C@H](O)[C@H]1O ZBDGHWFPLXXWRD-JGWLITMVSA-N 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 229950006780 n-acetylglucosamine Drugs 0.000 description 1
- 125000002347 octyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 230000020477 pH reduction Effects 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- OTYBMLCTZGSZBG-UHFFFAOYSA-L potassium sulfate Chemical class [K+].[K+].[O-]S([O-])(=O)=O OTYBMLCTZGSZBG-UHFFFAOYSA-L 0.000 description 1
- 235000011151 potassium sulphates Nutrition 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- MUPFEKGTMRGPLJ-ZQSKZDJDSA-N raffinose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O2)O)O1 MUPFEKGTMRGPLJ-ZQSKZDJDSA-N 0.000 description 1
- HEBKCHPVOIAQTA-ZXFHETKHSA-N ribitol Chemical compound OC[C@H](O)[C@H](O)[C@H](O)CO HEBKCHPVOIAQTA-ZXFHETKHSA-N 0.000 description 1
- NGFMICBWJRZIBI-UJPOAAIJSA-N salicin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=CC=C1CO NGFMICBWJRZIBI-UJPOAAIJSA-N 0.000 description 1
- 229940120668 salicin Drugs 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 238000004513 sizing Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 229940079827 sodium hydrogen sulfite Drugs 0.000 description 1
- 235000010267 sodium hydrogen sulphite Nutrition 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 210000005239 tubule Anatomy 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
- 229960003487 xylose Drugs 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses leuconostoc mesenteroides and application thereof in low-temperature silage. The Leuconostoc mesenteroides provided by the invention is specifically Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605, and the preservation number of the Leuconostoc mesenteroides in China center for type culture Collection is CCTCC NO: m2015254. The Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605CCTCC NO: m2015254 has stress resistance such as low temperature resistance, salt resistance, acid and alkali resistance, and can rapidly propagate and produce acid to reduce pH in the low temperature ensiling process (5 ℃), effectively inhibit the growth or production of harmful mixed bacteria, effectively retain nutrient components such as crude protein, crude fat, crude fiber, and the like, reduce non-nutrient substances such as crude ash components, and achieve the effect of long-term storage of the ensiling feed.
Description
Technical field
The invention belongs to microorganism field, it is related to one plant of Leuconostoc mesenteroides and its application in low temperature ensiling.
Background technology
Ensiling be under the anaerobic digestion of lactic acid bacteria by it is carbohydrate-modifying be organic acid so that pH reduction reach
To the purpose stored for a long time.Lactic acid bacteria and temperature are the deciding factors of ensilage fermentation quality quality.
Oat is that extremely frigid zones Winter-Spring raises the main forage crop mended, and because extremely frigid zones temperature low altitude area is high, is unfavorable for
The growth of lactic acid bacteria, it is difficult to the feed that is best in quality and can storing for a long time that ferments.
Therefore, the stronger bacterial strain of resistance of high-low temperature resistant, salt tolerant, acid and alkali-resistance is filtered out, and is added as leavening
It is added in low temperature ensiling oat feed, it will have huge application value to the silage making of extremely frigid zones.
The content of the invention
First purpose of the present invention is to provide one plant of Leuconostoc mesenteroides.
Leuconostoc mesenteroides provided by the present invention is specially Leuconostoc mesenteroides (Leuconostoc
Mesenteroides) QH1-605, the bacterial strain is preserved in China typical culture collection center (letter on April 28th, 2015
Claim CCTCC, address is：No. 299 Wuhan Universitys of Wuhan City, Hubei Province Wuchang District Bayi Road Wuhan University's collection in the school), its
Deposit number is CCTCC NO：M 2015254.
Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605 is from Qinghai Province's Qinghai Lake
Isolated on Bird Island wetland reed, the bacterial strain single bacterium colony is toroidal, and Gram's staining is positive, and does not produce hydrogen peroxide,
Unfermentable glucose aerogenesis, fermented type is homofermentation.Grown fine at 5 DEG C and 10 DEG C, more than 45 DEG C can not survive,
Show that the bacterial strain has good lower temperature resistance, heat-resisting quantity is poor；Grown fine in 3.0% NaCl culture medium, there is certain
Salt tolerance；Grown in pH3.5-9 culture environment well, show that acid-proof alkaline is preferable.Show the growth of the bacterial strain
Adaptability is very strong, is resistant to extreme environment Survival Reproduction.It is resistant to extreme environment Survival Reproduction.Its 16S rDNA sequences such as sequence table
Shown in middle sequence 1.
Second object of the present invention is to provide a kind of microbial inoculum.
The active component of microbial inoculum provided by the present invention is the Leuconostoc mesenteroides (Leuconostoc
mesenteroides)QH1-605CCTCC NO：M 2015254.
The microbial inoculum except comprising as activity into the Leuconostoc mesenteroides (Leuconostoc
mesenteroides)QH1-605CCTCC NO：Outside M 2015254, auxiliary material can be also included, such as (solvent is MRS solid mediums
Water, solute and its concentration are as follows：Peptone 10g/L, beef extract 10g/L, yeast extract 5g/L, glucose 20g/L, three acetate hydrates
Sodium 5g/L, Tween 80 1g/L, dipotassium hydrogen phosphate 2g/L, ammonium citrate 2g/L, magnesium sulfate 2g/L, manganese sulfate 0.25g/L, agar
17g/L)。
Third object of the present invention is to provide a kind of silage additive.
The active component of silage additive provided by the present invention is the Leuconostoc mesenteroides (Leuconostoc
mesenteroides)QH1-605CCTCC NO：M 2015254.
Fourth object of the present invention is to provide a kind of ensilage.
Contain the silage additive in ensilage provided by the present invention.
Leuconostoc mesenteroides (Leuconostoc mesenteroides) the QH1-605CCTCC NO：M 2015254
Or the microbial inoculum it is following it is any in application fall within protection scope of the present invention：
(a1) bacterium is suppressed；
(a2) bacterial inhibitor is prepared；
(a3) silage additive is prepared；
(a4) ensilage is prepared.
The application of (a1) is the application that non-diseases is diagnosed or treated.
In (a1) and (a2), the bacterium concretely micrococcus luteus and/or salmonella.
Wherein, Leuconostoc mesenteroides (Leuconostoc mesenteroides) the QH1-605CCTCC NO：
Suppression of the M2015254 or described microbial inoculums to the bacterium is embodied as the suppression under the conditions of 30 DEG C.
Application of the silage additive in the ensilage is prepared falls within protection scope of the present invention.
The 5th purpose of the present invention is to provide a kind of method for preparing the ensilage.
The method provided by the present invention for preparing the ensilage, specifically may include：By ensiling raw material and the goldbeater's skin
Leukonid (Leuconostoc mesenteroides) QH1-605CCTCC NO：M 2015254 is mixed, and carries out anaerobic solids
Fermentation, collects all tunnings, obtains the ensilage；
In the present invention, the ensiling raw material is oat complete stool；Specially milk stage oat complete stool, moisture
82.97%, cut into 1-2cm segments..
In the process, the oat complete stool and the Leuconostoc mesenteroides (Leuconostoc
mesenteroides)QH1-605CCTCC NO：M 2015254 proportioning can be 100g：105cfu；The fermentation is low temperature hair
Ferment；The low temperature can be 5 DEG C；The time of the fermentation can be 30 days.
Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605CCTCC NO provided by the present invention：
M 2015254 has the resistance such as low temperature resistant, salt tolerant, acid and alkali-resistance, and breeds rapidly during low temperature ensiling (5 DEG C) and produce acid
PH drops, effectively the growth or generation of the harmful miscellaneous bacteria of suppression, the nutritional ingredient such as thick protein, crude fat, crude fibre is effectively retained,
Non-nutritious matter such as coarse ash constituent reduction, reaches the long-term effect for preserving ensilage.
Preservation explanation
Strain name：Leuconostoc mesenteroides
Latin name：(Leuconostoc mesenteroides)
Strain number：QH1-605
Preservation mechanism：China typical culture collection center
Preservation mechanism is referred to as：CCTCC
Address：No. 299 Wuhan Universitys of Wuhan City, Hubei Province Wuchang District Bayi Road Wuhan University's collection in the school
Preservation date：On April 28th, 2015
Collection is registered on the books numbering：CCTCC NO：M 2015254
Embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
Material, reagent used etc., unless otherwise specified, are commercially obtained in following embodiments.
The separation and identification of embodiment 1, Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605
First, bacterial strain QH1-605 separation and screening
Qinghai Province Qinghai Lake Bird Island wetland reed 10g is taken, 90mL sterilized waters are put into, shaken 10 seconds, 1mL liquid is drawn
1.5mL centrifuge tubes are put in, 10 are diluted successively1, 103, 105Times, take the μ L of diluent liquid 20 to be respectively coated on MRS solid mediums,
30 DEG C of Anaerobic culturels 48 hours, take single bacterium colony MRS agar mediums to expand culture, are QH1- by the wherein one plant bacterium numbering of acquisition
605。
Wherein, the solvent of the MRS solid mediums is water, and solute and its concentration are as follows：Peptone 10g/L, beef extract
10g/L, yeast extract 5g/L, glucose 20g/L, sodium acetate trihydrate 5g/L, Tween 80 1g/L, dipotassium hydrogen phosphate 2g/L, lemon
Sour ammonium 2g/L, magnesium sulfate 2g/L, manganese sulfate 0.25g/L, agar 17g/L.
2nd, bacterial strain QH1-605 identification
Obtained bacterial strain QH1-605 is separated and screened to step one from the following aspects to identify.
1st, Morphological Identification
Step one is separated and screened after obtained bacterial strain QH1-605, Gram's staining, micro- Microscopic observation single bacterium colony shape
Rounded shape.
2nd, physiological and biochemical property is identified
It is Gram-positive that step one, which separates and screened obtained bacterial strain QH1-605, does not produce hydrogen peroxide, it is impossible to send out
Ferment glucose aerogenesis, fermented type is homofermentation.
Fermentation utilization powers of the bacterial strain QH1-605 to different carbon source is detected using API 50CH.As a result it is as shown in table 1.Can
See, the unfermentable utilization glycerine of bacterial strain QH1-605, D-R, D- xyloses, rhamnose, mannose, sorbierite, Alpha-Methyl-
D-MANNOSE, ursin, Alpha-Methyl-D- glucosides, D- turanoses, sucrose, melibiose, lactose, N-acetyl-glucosamine, pine three
Sugar, gossypose, starch, glycogen, 5- ketone groups-gluconate, D-Tag, L- trehaloses, gluconate, antierythrite, L- wood
Sugar, ribitol, Beta-methyl xyloside, galactitol, inosite, synanthrin, xylitol, D- lyxoses, D- trehaloses, D- is Arabic
Alcohol, L- arabites, fructose, sorbose, glucose, mannose, maltose, 2- ketone groups-gluconate does carbon source；It can ferment
Using ribose, L-arabinose, galactolipin, amarogentin, aesculin, cellobiose, salicin, trehalose is cooked carbon source；Can be with
It is faint to be fermented using gentiobiose as carbon source.
Fermentation utilization powers of the bacterial strain QH1-605 of table 1 to different carbon source
Note："+" represents positive, can utilize；"-" represents negative, i.e., can not utilize；" w " represents weakly positive, i.e., faint profit
With.
3rd, 16S rDNA sequence homology analysis
The bacterial strain QH1-605 of the gained of extraction step one genomic DNA, using it as template, is entered using bacterial universal primers
Performing PCR is expanded, and obtains bacterial strain QH1-605 16S rDNA fragments, and carries out sequencing, and its sequence is sequence 1 in sequence table.
Sequence 1 is subjected to BLAST (http in GenBank databases://blast.ncbi.nlm.nih.gov/Blast.cgi) it is same
Source property is compared, and determines strain classification.
According to above-mentioned form, analysis of physio biochemical characteristics and 16s rDNA sequence homology analysis results, by step one institute
The bacterial strain QH1-605 obtained is accredited as Leuconostoc mesenteroides (Leuconostoc mesenteroides), and April 28 in 2015
(abbreviation CCTCC, address is for day China typical culture collection center：Bayi Road No. 299 Wuhan in Wuhan City, Hubei Province Wuchang District are big
In school, Wuhan University's collection, postcode 430072), preserving number is CCTCC NO：M 2015254, its Classification And Nomenclature is intestines
Film leukonid (Leuconostoc mesenteroides) QH1-605.
Embodiment 2, Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605 different temperatures, pH and
Growth under salinity
1st, under different temperatures Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605 growth
By Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605CCTCC NO of activation：
M2015254 is inoculated in MRS fluid nutrient mediums, and the incubated 24h 5, at a temperature of 10,45 and 50 DEG C, uses spectrophotometer respectively
Absorbance value (OD600) at 600nm is determined, to observe Leuconostoc mesenteroides (Leuconostoc under different temperatures
Mesenteroides) QH1-605 growing state.
Wherein, the solvent of MRS fluid nutrient mediums is water, and solute and its concentration are as follows：Peptone 10g/L, beef extract 10g/
L, yeast extract 5g/L, glucose 20g/L, sodium acetate trihydrate 5g/L, Tween 80 1g/L, dipotassium hydrogen phosphate 2g/L, ammonium citrate
2g/L, magnesium sulfate 2g/L, manganese sulfate 0.25g/L；pH6.8.
As a result it is as shown in table 2, it is seen that Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605CCTCC
NO：M 2015254 grows fine at 5 DEG C and 10 DEG C, and more than 45 DEG C can not survive, and shows that the bacterial strain has good low temperature resistant
Property, heat-resisting quantity is poor.
2nd, under difference pH Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605 growth
The starting pH of MRS fluid nutrient mediums (formula is ibid) is distinguished with 1mol/L hydrochloric acid and 1mol/L sodium hydroxide
It is adjusted to 3.0,3.5,4.0,4.5,5.0,5.5,6.0,9.0 and 10.0,30 DEG C of constant temperature quiescent culture Leuconostoc mesenteroides
(Leuconostoc mesenteroides)QH1-605CCTCC NO：M 2015254, cultivates uncomfortable in 24h, incubation
Acid, with absorbance value (OD600) at spectrophotometric determination 600nm, to observe Leuconostoc mesenteroides under different pH
(Leuconostoc mesenteroides) QH1-605 growing state.
As a result it is as shown in table 2, it is seen that Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605CCTCC
NO：M 2015254 can grow well in pH3.5-9 culture environment, show that its acid-proof alkaline is preferable.
3rd, under different salinity Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605 growth
By Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605CCTCC NO of activation：
M2015254 be inoculated in respectively the weight/mass percentage composition containing NaCl be 3% and 6.5% MRS fluid nutrient mediums (formula ibid,
PH6.8), in 30 DEG C of constant temperature quiescent culture 24h, with absorbance value (OD600) at spectrophotometric determination 600nm, to observe not
With Leuconostoc mesenteroides under salinity (Leuconostoc mesenteroides) QH1-605CCTCC NO：M's 2015254
Growing state.
As a result it is as shown in table 2, it is seen that Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605CCTCC
NO：The well-growns under 3% NaCl concentration of M 2015254, show that it has certain salt tolerance.
4th, (5 DEG C) culture results of regular determination pH changes of low temperature
MRS fluid nutrient mediums (are formulated ibid, pH6.8) 1.5ml and are put in 2ml centrifuge tubes, QH1-666 single bacterium colonies, 5 is accessed
DEG C culture, respectively at 7 days, 10 days, at 14 days with pH acidometers determine filtrate pH value.Experiment is repeated 3 times, as a result with average value
Form represent.
As a result as shown in table 2, it is seen that (5 DEG C) of cryogenic conditions are cultivated 7 days, the pH of fermentation system drops to 4.5, culture 10 days and
After 14 days, pH falls below 4 and 3.5 reduced levels respectively, shows Leuconostoc mesenteroides (Leuconostoc
mesenteroides)QH1-605CCTCC NO：M 2015254 can effectively reduce pH.
The growth characteristics of the bacterial strain QH1-605 of table 2 under various circumstances
Note："+" represents that (OD600 is considered as well-grown to well-grown more than 0.3, and it is growth that 0.3 is less than or equal to more than 0.2
It is faint).
According to result above, it is known that Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605CCTCC
NO：M 2015254 growth adaptation is very capable, is resistant to extreme environment Survival Reproduction.
The Antibacterial Activity of embodiment 3, Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605
For examination bacterium：Micrococcus luteus, micrococcus luteus, salmonella and Escherichia coli.
1st, by Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605CCTCC NO of activation：
M2015254 is inoculated in MRS fluid nutrient mediums (formula ibid, pH6.8), 30 DEG C of culture 48h, zymotic fluid through 10000rpm from
Heart 5min, takes supernatant to be used to determine.
2nd, its bacteriostatic activity is determined using Oxford cup double-layer agar technique, sterilized agar medium is heated to completely to melt
Change, be poured in culture dish, per ware 15ml (lower floor), treat that it solidifies.In addition, the PDA culture medium of thawing is cooled into 50 DEG C or so
It is mixed into for examination bacterium, the culture medium 5ml for being mixed with bacterium is added to (upper strata) to be solidified on the culture medium solidified.Existed with sterile working
Media surface directly vertically puts Oxford cup (internal diameter 6mm, external diameter 8mm, high 10mm circular tubule), gently pressurizes, makes it
Tight is contacted with culture medium, the Leuconostoc mesenteroides (Leuconostoc that step 1 is obtained is added in cup
mesenteroides)QH1-605CCTCC NO：The zymotic fluids of M 2015254.Fill it up with rearmounted 37 DEG C to cultivate 16-18 hours, observation
As a result, inhibition zone dipstick metering.Control is used as using MRS fluid nutrient mediums substitution zymotic fluid.Experiment is repeated 3 times, results averaged.
It is as a result as shown in table 3, it is seen that：Under 30 DEG C of condition of culture, Leuconostoc mesenteroides (Leuconostoc
mesenteroides)QH1-605CCTCC NO：M 2015254 has to micrococcus luteus and salmonella preferably to be suppressed to make
With.
The bacterial strain QH1-605 of table 3 bacteriostatic activity
Note：Antibacterial circle diameter includes Oxford cup external diameter (7.8mm)；"-" indicates no inhibition zone.
Embodiment 4, Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605 ensiling oats
Ensiling raw material：Milk stage oat complete stool, moisture 82.97% cuts into 1-2cm segments.
First, oat ensilage is prepared
1st, with Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605CCTCC NO：M 2015254
Oat ensilage is prepared as feed addictive
(1) Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605CCTCC NO：The bacterium of M 2015254
The preparation of suspension
Under aseptic condition, Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605CCTCC NO are taken：M
2015254 are inoculated in 30 DEG C of overnight incubations in MRS fluid nutrient mediums, obtain Leuconostoc mesenteroides (Leuconostoc
mesenteroides)QH1-605CCTCC NO：Leuconostoc mesenteroides in the bacteria suspensions of M 2015254, bacteria suspension
(Leuconostoc mesenteroides)QH1-605CCTCC NO：M 2015254 content is 106cfu/ml。
Wherein, the solvent of MRS fluid nutrient mediums is water, and solute and its concentration are as follows：Peptone 10g/L, beef extract 10g/
L, yeast extract 5g/L, glucose 20g/L, sodium acetate trihydrate 5g/L, Tween 80 1g/L, dipotassium hydrogen phosphate 2g/L, ammonium citrate
2g/L, magnesium sulfate 2g/L, manganese sulfate 0.25g/L；pH6.8.
(2) with Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605CCTCC NO：M2015254
Oat ensilage is prepared as feed addictive
1-2cm or so length (moisture 82.97%) is cut into chopper after milk stage oat complete stool is gathered in, 100g is taken
It is put into bag silo, Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1- that step (1) is obtained
605CCTCC NO：The μ l of 2015254 every bag of bacteria suspensions of M 100 are separately added into bag silo, are mixed, are taken out very using vacuum packing machine
Seal, be put into 5 DEG C of refrigerator-freezers after sky.
2nd, oat ensilage is compareed
1-2cm or so length (moisture 82.97%) is cut into chopper after milk stage oat complete stool is gathered in, 100g is taken
It is put into bag silo, every bag of 100 μ l of sterilized water is separately added into bag silo, mixes, vacuumized using vacuum packing machine rear close
Envelope, is put into 5 DEG C of refrigerator-freezers.
2nd, the measure that oat pH changes in low temperature ensilage
Feed Sample 10g is added in 90ml sterile distilled waters, 30s is shaken using vortex oscillator, takes liquid to pass through
Filter paper is filtered, and the pH value of filtrate is determined using pH acidometers.Experiment is repeated 3 times, as a result the table in the form of mean+SD
Show.
It is as a result as shown in table 4, it is seen that：Under the conditions of low temperature (5 DEG C) ensiling, Leuconostoc mesenteroides is added in ensiling the 1st day
(Leuconostoc mesenteroides)QH1-605CCTCC NO：M 2015254 oat ensilage pH is substantially less than
Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605CCTCC NO are not added：M 2015254 pair
According to group (P≤0.05)；In ensiling the 3rd day, the 7th day, the 30th day, Leuconostoc mesenteroides (Leuconostoc
mesenteroides)QH1-605CCTCC NO：The addition group ensiling oat pH of M 2015254 distinguish pole and are substantially less than control group (P
≤0.01)；In ensiling the 30th day, pH fell below 4.12, the level preserved for a long time beneficial to ensilage.Result above shows, 5
Under DEG C low temperature environment, Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605CCTCC NO：M 2015254
Effectively ensiling oat pH can be reduced to beneficial to ensiling level rapidly
Oat pH changes during the low temperature ensiling of table 4 (5 DEG C)a
| 1 day | 3 days | 7 days | 30 days | |
| Control | 6.71±0.08 | 6.67±0.02 | 6.35±0.39 | 6.06±0.06 |
| Bacterial strain QH1-605 | 6.52±0.07 | 6.19±0.1 | 4.36±0.05 | 4.12±0.19 |
| Standard error | 0.05 | 0.11 | 0.1 | 0.07 |
| Conspicuousness | * | ** | ** | ** |
Note：aMean+SD (n=3)." NS " represents that difference is not notable, P>0.05；" * " represents significant difference
(P≤0.05)；" * * " represent pole significant difference (P≤0.01)
3rd, the microbiology turbidity of oat is determined in low temperature ensilage
Using plate dilution assay method microbiology turbidity.Feed Sample 10g is added in 90ml sterile distilled waters, utilized
Vortex oscillator shakes 30s, and then 10 times of gradient dilutions take 10 respectively-1、10-3With 10-5Each 20 μ l coatings of times sample diluting liquid
(it is used to detect saccharomycete and mould, according to bacterium colony in MRS (being used to detect lactic acid bacteria), BLB (being used to detect Escherichia coli), PDA
Form, naked eyes are distinguished, and mold colony is in villiform, and the color of its spore is presented in cotton-shaped, spider reticulation, and saccharomycete is smaller,
Shaft-like, helical form is spherical, and bacterium colony surface is smooth sticky or dry) NA (for detecting aerobic bacteria) culture medium；By 10-1
With 10-2Times sample diluting liquid 1ml is after 75 DEG C of water-bath water-bath 15min, and taking 20 μ l to be coated on NA respectively (is used to detect gemma bar
Bacterium) and CLO (for detecting clostridium) culture medium (formula of CLO culture mediums：Contain peptone 15g, soybean egg in every liter of culture medium
White peptone 7.5g, yeast extract 7.5g, beef extract 7.5g, ferric citrate 1g, sodium hydrogensulfite 1g, L-cysteine hydrochloride
0.75g, agar 15g, surplus is water) on, these coated culture mediums are inverted and are put in 30 DEG C of constant incubator culture 48h,
Wherein MRS and CLO culture mediums should be positioned over anaerobic culture box, and other culture mediums are positioned over common constant incubator.Culture
48 hours postscript single bacterium colony numbers are n, bacterium colony (logCFU/g)=log10[(n×10)/20×10-3].Experiment is repeated 3 times, knot
Fruit is represented in the form of mean+SD.
It is as a result as shown in table 5, it is seen that：Ensiling the 1st day and the 3rd day, addition Leuconostoc mesenteroides (Leuconostoc
mesenteroides)QH1-605CCTCC NO：M 2015254 ensiling oat group lactic acid bacterium number pole is significantly higher than control group
Level (P≤0.01)；Ensiling the 3rd day, compared with the control, Leuconostoc mesenteroides (Leuconostoc mesenteroides)
QH1-605CCTCC NO：The addition groups of M 2015254 do not detect saccharomycete and clostridium, Leuconostoc mesenteroides (Leuconostoc
mesenteroides)QH1-605CCTCC NO：The growth vigors of M 2015254 substantially, may effectively inhibit saccharomycete and
The generation of clostridium.Ensiling the 30th day, Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605CCTCC NO：
The addition groups of M 2015254 compared with the control, do not detect Escherichia coli.In a word, Leuconostoc mesenteroides is added in ensilage
(Leuconostoc mesenteroides)QH1-605CCTCC NO：M 2015254, Leuconostoc mesenteroides (Leuconostoc
mesenteroides)QH1-605CCTCC NO：M 2015254 is mushroomed out as dominant strain.Consolidated statement 3, addition goldbeater's skin is bright
Beading bacterium (Leuconostoc mesenteroides) QH1-605CCTCC NO：M 2015254 substantially reduces ensilage pH,
Effectively suppress Escherichia coli, bacillus, saccharomycete, the growth of harmful miscellaneous bacteria such as clostridium.
The microbiology turbidity (logCFU/g) of oat during the low temperature ensiling of table 5 (5 DEG C)a
Note：aMean+SD (n=3).Same file contain different capitalizations represent otherness extremely significantly (P≤
0.01)；Same file contains different lowercase letter indication differences significantly (P≤0.05).ND represents not detect.
4th, the measure that the free water of oat, dry weight and crude ash content change in low temperature ensilage
1st, free water content and the assay method of dry weight content
65 DEG C, a blank sheet of paper is ordered into a paper bag first, paper bag is weighed on assay balance by 48h seasonings,
W1 is designated as, Feed Sample is put into paper bag, claims gross weight, W2 is designated as, constant temperature is divulged information after 65 DEG C of dry 48h, is positioned over drier
Weighed after middle 30min, be designated as W3.Free moisture (%)=(W2-W3)/(W2-W1) × 100.Dry weight (%)=100%- dissociates
Moisture (%).Experiment is repeated 3 times, and is as a result represented in the form of mean+SD.
2nd, the measure of coarse ash (Crude ash)
(1) porcelain crucible is heated after 2h in 530 DEG C of Muffle furnaces, and taking-up is positioned over 1h in drier and cooled, and weighs, is designated as
W1。
(2) 2g (W2) left and right Feed Sample is weighed in porcelain crucible by assay balance, is positioned on electric furnace and is ashed,
Untill cigarette disperses, take out porcelain crucible with crucible tongs and be positioned in 530 DEG C of Muffle furnaces after heating 2h, taking-up is positioned over drier
Middle 1h is cooled, and is weighed, and is designated as W3.
Coarse ash (%)=(W3-W1)/W2 × 100.
Experiment is repeated 3 times, and is as a result represented in the form of mean+SD.
It is as a result as shown in table 6, it is seen that：In whole ensilage, Leuconostoc mesenteroides (Leuconostoc is added
mesenteroides)QH1-605CCTCC NO：M 2015254 can reduce the percent water of ensilage, and increase is blue or green
The dry weight percentage content of feed is store, crude ash content can also be reduced, and in ensiling the 3rd day, adds Leuconostoc mesenteroides
(Leuconostoc mesenteroides)QH1-605CCTCC NO：M2015254 feed, its coarse ash and free water contain
Amount is significantly reduced (P≤0.05) compared with the control, and dry weight content is dramatically increased (P≤0.05).Show 5 DEG C of low temperature ensilages
In, Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605CCTCC NO：M 2015254 can be effective
The content of non-nutritious matter coarse ash is reduced, cost of transportation is reduced, but nutriment dry weight is then slightly increased.
The free water of oat, dry weight and crude ash content change (%/fresh weight) during the low temperature ensiling of table 6 (5 DEG C)a
Note：aMean+SD (n=3)；" * " represents significant difference (P≤0.05)；" NS " represents that difference is not notable
(p ＞ 0.05).
5th, the crude fat of oat in low temperature ensilage, crude protein, neutral detergent fiber and acid detergent fiber change
Determine
1st, the measure of crude fat
Using aether extraction.Feed Sample 3h extractive distillations crude fat in ether, in addition to neutral fat, phosphate ester
Matter, free fatty, fat-soluble pigment etc. are also included.
(1) fatty bottle, which is positioned in 100 DEG C of baking ovens, dries after 2h, and taking-up, which is placed in drier, cools 1h, weighs, is designated as
W1。
(2) assay balance weighs 1g (W2) left and right Feed Sample and is placed on filter paper, fills in filter paper in fatty bucket after wrapping,
Blocked up with absorbent cotton.
(3) the fatty bucket that will be equipped with Feed Sample is put into fatty withdrawing device, connects fatty bottle, adds about 50ml ether
In fatty bottle bottle, cooling water system is opened, starts more than 3h extractings.
(4) complete after extracting, take out fatty bucket, fatty bottle is placed in 100 DEG C of baking ovens after 3h dryings, taking-up is placed on dry
30m is cooled down in dry device, is weighed, W3 is designated as.
Crude fat (%)=(W3-W1)/W2 × 100.Experiment is repeated 3 times, as a result the table in the form of mean+SD
Show.
2nd, the measure of crude protein
Sizing technique is boiled using disappearing and determines crude protein.
(1) about 1g (W1) Feed Sample is weighed by assay balance, pan paper is put into disappear after wrapping and boiled in pipe, added
0.2g copper sulphate and 6.0g potassium sulfates.
(2) the 20ml concentrated sulfuric acids are gently added to boil in pipe in disappearing, gently shakes, sample is fully distributed in concentrated sulfuric acid.
(3) it will disappear to boil pipe connection and be positioned over to disappear and boil on pipe, and prevent the volatilization of sulfuric acid.
(4) circulating water device is opened, will disappear to boil pipe and be placed on to disappear and boil on device, begin heat to 420 DEG C.
(5) wait disappearing and boil liquid in pipe and be changed into after green transparent, continue 420 DEG C and disappear to boil 2h.
(6) disappear and boil after decomposition completes, turn off to disappear and boil device, remove to disappear to boil pipe and be put on thermal insulation board and cool.
(7) after cooling down, digest tube is put into kjeldahl apparatus, starts titration (according to AOAC (Official Methods
of Analysis[M].15th ed.Association of official analytical chemists.Arlington,
VA.1999) method described is analyzed).
Total nitrogen content (%)=n (V1-V2) × M × 0.014/W1 × 100；
Crude protein quality (%)=total nitrogen content (%) × 6.25；
Wherein, V1：Titer ml；V2：Blank titration amount；M：Titrating solution HCl concentration；W1：Example weight.
Experiment is repeated 3 times, and is as a result represented in the form of mean+SD.
3rd, the measure of neutral detergent fiber (neutral detergent fiber, NDF)
After plant feed boils processing through neutral detergent, undissolved residue is neutral detergent fiber, predominantly
Cell wall constituent, neutral detergent fiber includes cellulose, this quality and hemicellulose, may be used as quantitative Livestock roughage
Feed intake.
(1) 20min opens crude fiber test instrument device and cooling circulating water in advance.
(2) assay balance weighs about 0.5g (W1) left and right Feed Sample and is positioned in glass pot.
(3) machine on, adds (3g/100ml) lauryl sodium sulfate of 100ml neutral detergents 3% and appropriate octyl alconyl is made
Defoamer, 100 DEG C are heated to boiling, keep 50 DEG C or so heating 70min.
(4) glass pot after suction filtration, is removed, acetone rinsing three times is utilized.
(5) glass pot is put in fume hood and be dried overnight.
(6) later glass pot will be dried to be positioned in baking oven, more than 135 DEG C of constant temperature aeration-drying 2h are put into drying
Cooled down in device after 1h, weigh, be designated as W2.
(7) glass pot is put on electric furnace and is ashed, until cigarette disperses, taking out crucible using crucible tongs is positioned over 530
In DEG C Muffle furnace after heating ashing 2h, taking-up is positioned over 1h in drier and cooled, and weighs, is designated as W3.
NDF (%)=(W2-W3)/W1 × 100.
Experiment is repeated 3 times, and is as a result represented in the form of mean+SD.
4th, the measure of acid detergent fiber (acid detergent fiber, ADF)
Acid detergent fiber assay method：After being handled using acid detergent, residual residue is acid detergent fiber,
Including lignin and cellulose.Residue of the acid detergent fiber after sulfuric acid treatment is lignin, fine from acidic cleaning
The residue that 72% sulfuric acid treatment is subtracted in dimension value is feed fibre cellulose content.
(1) 20min opens crude fiber test instrument device and cooling circulating water in advance.
(2) assay balance weighs about 0.5g (W1) left and right Feed Sample and is positioned in glass pot.
(3) machine on, adds (2g/100ml) cetyl trimethylammonium bromide of 100ml acid detergents 2% and appropriate fourth
Octanol makees defoamer, and 100 DEG C are heated to boiling, keeps 50 DEG C or so heating 70min.
(4) glass pot after suction filtration, is removed, acetone rinsing three times is utilized.
(5) glass pot is put in fume hood and be dried overnight.
(6) later glass pot will be dried to be positioned in baking oven, more than 135 DEG C of constant temperature aeration-drying 2h are put into drying
Cooled down in device after 1h, weigh, be designated as W2.
(7) glass pot is put on electric furnace and is ashed, until cigarette disperses, taking out crucible using crucible tongs is positioned over 530
In DEG C Muffle furnace after heating ashing 2h, taking-up is positioned over 1h in drier and cooled, and weighs, is designated as W3.
ADF (%)=(W2-W3)/W1 × 100.
Experiment is repeated 3 times, and is as a result represented in the form of mean+SD.
It is as a result as shown in table 7, it is seen that：Under the conditions of low temperature (5 DEG C) ensiling, Leuconostoc mesenteroides (Leuconostoc
mesenteroides)QH1-605CCTCC NO：Acidic cleaning of the addition groups of M 2015254 compared with control group to ensilage
Fiber content has no significant effect；Ensiling the 3rd day, Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-
605CCTCC NO：The neutral detergent fiber content of the addition groups of M 2015254 is substantially less than control group (P<0.05), in addition, neutral
Content is on a declining curve with the lengthening of period of storage, control group and Leuconostoc mesenteroides (Leuconostoc
mesenteroides)QH1-605CCTCC NO：The addition group neutral detergent fiber contents of M 2015254 are respectively from 56.93% He
57.27% falls below 52.00% and 52.72%；Compared with control group, Leuconostoc mesenteroides (Leuconostoc is added
mesenteroides)QH1-605CCTCC NO：M 2015254 does not show to the crude protein and crude fat content of ensiling oat
Influence (P ＞ 0.05) is write, in 5 DEG C of low temperature ensilages, crude fat and crude protein composition retain preferable.
Crude fat, crude protein, neutral detergent fiber and the acid detergent fiber of oat during the low temperature ensiling of table 7 (5 DEG C)
Change (%/fresh weight)a
Note：aMean+SD (n=3)；" NS " represents difference not significantly (p ＞ 0.05)；" * " represents significant difference
(p≤0.05).NDF is neutral detergent fiber；ADF is acid detergent fiber.
In summary, it is seen that Leuconostoc mesenteroides (Leuconostoc mesenteroides) QH1-605CCTCC NO：M
2015254 breed rapidly during low temperature ensiling (5 DEG C) and produce acid drop pH, the effective growth or production for suppressing to be harmful to miscellaneous bacteria
Raw, the nutritional ingredient such as crude protein, crude fat, crude fibre is effectively retained, non-nutritious matter such as coarse ash constituent reduction, is reached long-term
Preserve the effect of ensilage.
Claims (9)
1. Leuconostoc mesenteroides（Leuconostoc mesenteroides）QH1-605, it is in China typical culture collection
The deposit number of the heart is CCTCC NO：M 2015254.
2. a kind of microbial inoculum, it is characterised in that：The active component of the microbial inoculum is the Leuconostoc mesenteroides described in claim 1
（Leuconostoc mesenteroides）QH1-605.
3. a kind of silage additive, it is characterised in that：The active component of the silage additive is claim 1 institute
The Leuconostoc mesenteroides stated（Leuconostoc mesenteroides）QH1-605.
4. a kind of ensilage, it is characterised in that：Added in the ensilage containing the ensilage described in claim 3
Agent.
5. ensilage according to claim 4, it is characterised in that：The ensilage is according to comprising the following steps
What method was prepared：
By the Leuconostoc mesenteroides described in ensiling raw material and claim 1（Leuconostoc mesenteroides）QH1-605
Mixing, carries out solid anaerobic fermentation, collects all tunnings, obtains the ensilage；
The ensiling raw material is specially oat complete stool；
The oat complete stool and the Leuconostoc mesenteroides（Leuconostoc mesenteroides）QH1-605 proportioning is
100g：105cfu；
The fermentation is cold fermentation；The low temperature is 5 DEG C；
The time of the fermentation is 30 days.
6. the Leuconostoc mesenteroides described in claim 1（Leuconostoc mesenteroides）QH1-605 or claim
Microbial inoculum described in 2 it is following it is any in application：
（a1）Prepare bacterial inhibitor；
（a2）Prepare the silage additive described in claim 3；
（a3）Prepare the ensilage described in claim 4；
Described（a1）In, the bacterium is specially micrococcus luteus or salmonella.
7. application according to claim 6, it is characterised in that：The Leuconostoc mesenteroides（Leuconostoc
mesenteroides）The suppression that is suppressed to 30 DEG C under the conditions of of the QH1-605 or described microbial inoculums to the bacterium.
8. application of the silage additive in the ensilage described in claim 4 or 5 is prepared described in claim 3.
9. the method for ensilage described in claim 4 is prepared, including：Ensiling raw material and goldbeater's skin described in claim 1 is bright
Beading bacterium（Leuconostoc mesenteroides）QH1-605 is mixed, and carries out solid anaerobic fermentation, collects all fermentation productions
Thing, obtains the ensilage；
The ensiling raw material is specially oat complete stool；
The oat complete stool and the Leuconostoc mesenteroides（Leuconostoc mesenteroides）QH1-605 proportioning is
100g：105cfu；
The fermentation is cold fermentation；The low temperature is 5 DEG C；
The time of the fermentation is 30 days.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510340177.0A CN104894031B (en) | 2015-06-18 | 2015-06-18 | Leuconostoc mesenteroides and application thereof in low-temperature silage |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510340177.0A CN104894031B (en) | 2015-06-18 | 2015-06-18 | Leuconostoc mesenteroides and application thereof in low-temperature silage |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104894031A CN104894031A (en) | 2015-09-09 |
| CN104894031B true CN104894031B (en) | 2017-09-26 |
Family
ID=54026992
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510340177.0A Expired - Fee Related CN104894031B (en) | 2015-06-18 | 2015-06-18 | Leuconostoc mesenteroides and application thereof in low-temperature silage |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104894031B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110177866B (en) * | 2016-07-15 | 2023-06-20 | Cj第一制糖株式会社 | Leuconostoc mesenteroides CJLM627 strain with low gas production and method for preparing pickle by using strain |
| RU2731642C2 (en) * | 2016-07-15 | 2020-09-07 | СиДжей ЧЕИЛДЗЕДАНГ КОРПОРЕЙШН | Cjlm119 leuconostoc mesenteroides strain, which produces reduced amount of gas, and method for preparing kimchi using said strain |
| HUE065092T2 (en) | 2016-07-15 | 2024-04-28 | Cj Cheiljedang Corp | Leuconostoc mesenteroides cjlm181 strain producing reduced amount of gas, and kimchi production method using same |
| CN118028165B (en) * | 2024-02-29 | 2024-07-09 | 江苏省农业科学院 | Leuconostoc mesenteroides and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101974450A (en) * | 2010-09-13 | 2011-02-16 | 郑州大学 | Leuconostoc mesenteroides and application thereof |
| CN102643767A (en) * | 2012-04-19 | 2012-08-22 | 郑州大学 | Lactobacillus plantarum and application thereof in fermenting and ensiling sweet potato stem and leaf |
| CN104073454A (en) * | 2014-06-18 | 2014-10-01 | 兰州大学 | Low-temperature-resistant lactobacillus planetarium and application thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| MX2014010020A (en) * | 2012-02-21 | 2014-12-05 | Dupont Nutrition Biosci Aps | Composition comprising fermentation products from bacillus subtilis. |
-
2015
- 2015-06-18 CN CN201510340177.0A patent/CN104894031B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101974450A (en) * | 2010-09-13 | 2011-02-16 | 郑州大学 | Leuconostoc mesenteroides and application thereof |
| CN102643767A (en) * | 2012-04-19 | 2012-08-22 | 郑州大学 | Lactobacillus plantarum and application thereof in fermenting and ensiling sweet potato stem and leaf |
| CN104073454A (en) * | 2014-06-18 | 2014-10-01 | 兰州大学 | Low-temperature-resistant lactobacillus planetarium and application thereof |
Non-Patent Citations (7)
| Title |
|---|
| Identification and Characterization of Leuconostoc gelidum,Isolated from Kimchi, a Fermented Cabbage Product;Bong-Joon Kim等;《The Journal of Microbiology》;20000930;第38卷(第3期);第132-136页 * |
| Phenotypic and Phylogenetic Analysis of Lactic Acid Bacteria Isolated from Forage Crops and Grasses in the Tibetan Plateau;Huili Pang等;《The Journal of Microbiology》;20121231;第50卷(第1期);摘要，第63-71页 * |
| 一株产广谱细菌素乳酸菌菌株的筛选及鉴定;吕好新;《中国乳品工业》;20131231;第41卷(第10期);第8-10页 * |
| 耐低温乳酸菌的分离与优化培养;储徐建等;《草业科学》;20170731;第31卷(第7期);第1380-1388页 * |
| 膜明串珠茵的研究和应用进展;李文斌等;《食品工程》;20061231(第4期);第3-5页 * |
| 西藏地区传统发酵乳中乳酸菌多样性及微生物数量分析;陈芝兰等;《生物工程》;20131231;第34卷(第17期);第140-145页 * |
| 高抗菌活性乳酸菌的筛选及菌种鉴定;李东霞等;《饲料工业》;20131231;第34卷(第19期);第13-17页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104894031A (en) | 2015-09-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104946564B (en) | One lactobacillus plantarum and its application in low temperature ensiling | |
| CN104894030B (en) | One plant of Lactococcus lactis subsp. lactis and its application in low temperature ensiling | |
| CN103205374B (en) | Lactic acid bacterium capable of reducing aflatoxin B1 in fermented feed and application of lactic acid bacterium | |
| CN102643767B (en) | Lactobacillus plantarum and application thereof in fermenting and ensiling sweet potato stem and leaf | |
| CN104894031B (en) | Leuconostoc mesenteroides and application thereof in low-temperature silage | |
| CN109161498B (en) | Bacillus subtilis M406 and application thereof in preparation of bacteriocin and cellulase | |
| CN104894029B (en) | Leuconostoc mesenteroides and its application in low temperature ensiling | |
| CN104320980B (en) | Protective foods containing pediococcus acidilactici J9 and its preparation method | |
| CN112662599B (en) | Poultry source Bacillus belgii CL-4 and application thereof | |
| CN107217018A (en) | Resistance bacillus subtilis and its application | |
| CN103952336B (en) | Bacillus licheniformis, bacillus subtilis and lactobacillus casei preparation and preparation | |
| CN103952324A (en) | Kluyveromyces marxianus C2 and preparation method and application thereof | |
| CN111304119A (en) | Feeding bacillus subtilis for degrading fumonisins and application thereof | |
| CN117070427B (en) | Lactobacillus buchneri and silage starter thereof | |
| CN104911131B (en) | Enterococcus mundtii and application thereof in low-temperature silage | |
| CN106479944A (en) | A kind of bacillus amyloliquefaciens and its application | |
| CN104087529B (en) | bacillus pumilus (Bacillus pumilus) JY2 and application thereof | |
| CN113151032A (en) | Bacillus subtilis with efficient gossypol degradation capability and application thereof | |
| CN111748490A (en) | Lactobacillus sake and application thereof | |
| CN115011515B (en) | Strain for increasing plant alcohol content in alfalfa silage and application thereof | |
| CN113999801B (en) | Bacillus belicus capable of modifying wheat bran and application thereof | |
| CN110452843A (en) | A kind of feeding bacillus subtilis and its application | |
| CN113897312B (en) | Preparation and application of animal feeding microbial inoculum | |
| CN114854629A (en) | Lactobacillus plantarum C-37 and application thereof in fermenting corn germ meal | |
| CN102640886B (en) | Applications of Lactobacillus plantarum in Brassica oleracea outer wrapper leaf silage |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170926 Termination date: 20190618 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |