CN115702667A - Method for ensiling sesbania and sweet sorghum mixture - Google Patents
Method for ensiling sesbania and sweet sorghum mixture Download PDFInfo
- Publication number
- CN115702667A CN115702667A CN202110907045.7A CN202110907045A CN115702667A CN 115702667 A CN115702667 A CN 115702667A CN 202110907045 A CN202110907045 A CN 202110907045A CN 115702667 A CN115702667 A CN 115702667A
- Authority
- CN
- China
- Prior art keywords
- lactobacillus
- sesbania
- silage
- buchneri
- fermentation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 235000011684 Sorghum saccharatum Nutrition 0.000 title claims abstract description 51
- 244000138286 Sorghum saccharatum Species 0.000 title claims abstract description 49
- 238000000034 method Methods 0.000 title claims abstract description 29
- 239000000203 mixture Substances 0.000 title claims abstract description 23
- 244000275012 Sesbania cannabina Species 0.000 title description 54
- 239000004460 silage Substances 0.000 claims abstract description 90
- 241000186679 Lactobacillus buchneri Species 0.000 claims abstract description 59
- 238000000855 fermentation Methods 0.000 claims abstract description 55
- 230000004151 fermentation Effects 0.000 claims abstract description 52
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 38
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 38
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 38
- 241000186685 Lactobacillus hilgardii Species 0.000 claims abstract description 36
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims abstract description 32
- 238000002360 preparation method Methods 0.000 claims abstract description 26
- 241000186660 Lactobacillus Species 0.000 claims abstract description 20
- 229940039696 lactobacillus Drugs 0.000 claims abstract description 20
- 241000219782 Sesbania Species 0.000 claims abstract description 16
- 239000004310 lactic acid Substances 0.000 claims abstract description 16
- 235000014655 lactic acid Nutrition 0.000 claims abstract description 16
- 241000894006 Bacteria Species 0.000 claims abstract description 15
- 150000001875 compounds Chemical class 0.000 claims abstract description 13
- 238000002156 mixing Methods 0.000 claims abstract description 11
- 239000002994 raw material Substances 0.000 claims abstract description 11
- 239000007787 solid Substances 0.000 claims abstract description 8
- 239000004480 active ingredient Substances 0.000 claims abstract description 5
- 239000002131 composite material Substances 0.000 claims description 18
- 239000002068 microbial inoculum Substances 0.000 claims description 9
- 238000004321 preservation Methods 0.000 claims description 8
- 238000009629 microbiological culture Methods 0.000 claims description 4
- 239000000463 material Substances 0.000 claims description 3
- 241001465754 Metazoa Species 0.000 claims description 2
- 241000196324 Embryophyta Species 0.000 abstract description 11
- 230000000813 microbial effect Effects 0.000 abstract description 11
- 210000002421 cell wall Anatomy 0.000 abstract description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 24
- 230000001580 bacterial effect Effects 0.000 description 23
- 239000003795 chemical substances by application Substances 0.000 description 16
- 239000000047 product Substances 0.000 description 13
- 239000001963 growth medium Substances 0.000 description 10
- DNIAPMSPPWPWGF-GSVOUGTGSA-N (R)-(-)-Propylene glycol Chemical compound C[C@@H](O)CO DNIAPMSPPWPWGF-GSVOUGTGSA-N 0.000 description 8
- 241001052560 Thallis Species 0.000 description 8
- 235000011054 acetic acid Nutrition 0.000 description 8
- DNIAPMSPPWPWGF-UHFFFAOYSA-N monopropylene glycol Natural products CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 8
- 229960004063 propylene glycol Drugs 0.000 description 8
- 235000013772 propylene glycol Nutrition 0.000 description 8
- 230000000694 effects Effects 0.000 description 7
- 235000019750 Crude protein Nutrition 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- SRBFZHDQGSBBOR-SOOFDHNKSA-N D-ribopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@@H]1O SRBFZHDQGSBBOR-SOOFDHNKSA-N 0.000 description 5
- 108020004414 DNA Proteins 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- 238000009630 liquid culture Methods 0.000 description 5
- 244000005700 microbiome Species 0.000 description 5
- -1 polyethylene Polymers 0.000 description 5
- 241000894007 species Species 0.000 description 5
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 4
- 229930006000 Sucrose Natural products 0.000 description 4
- 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 4
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 4
- 229910052799 carbon Inorganic materials 0.000 description 4
- 239000011248 coating agent Substances 0.000 description 4
- 238000000576 coating method Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000002504 physiological saline solution Substances 0.000 description 4
- 238000012163 sequencing technique Methods 0.000 description 4
- 239000005720 sucrose Substances 0.000 description 4
- 244000063299 Bacillus subtilis Species 0.000 description 3
- 235000014469 Bacillus subtilis Nutrition 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- PYMYPHUHKUWMLA-LMVFSUKVSA-N Ribose Natural products OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 description 3
- HMFHBZSHGGEWLO-UHFFFAOYSA-N alpha-D-Furanose-Ribose Natural products OCC1OC(O)C(O)C1O HMFHBZSHGGEWLO-UHFFFAOYSA-N 0.000 description 3
- 230000025938 carbohydrate utilization Effects 0.000 description 3
- 239000002689 soil Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- 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 2
- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
- OBMBUODDCOAJQP-UHFFFAOYSA-N 2-chloro-4-phenylquinoline Chemical compound C=12C=CC=CC2=NC(Cl)=CC=1C1=CC=CC=C1 OBMBUODDCOAJQP-UHFFFAOYSA-N 0.000 description 2
- 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 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-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- 229930091371 Fructose Natural products 0.000 description 2
- 239000005715 Fructose Substances 0.000 description 2
- 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 2
- 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 2
- 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 2
- 239000004698 Polyethylene Substances 0.000 description 2
- 240000006394 Sorghum bicolor Species 0.000 description 2
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 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 2
- 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 2
- 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 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000000835 fiber Substances 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 229930182830 galactose Natural products 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 238000012165 high-throughput sequencing Methods 0.000 description 2
- 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 2
- 238000002386 leaching Methods 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
- 235000016709 nutrition Nutrition 0.000 description 2
- 229920000573 polyethylene Polymers 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 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
- OMDQUFIYNPYJFM-XKDAHURESA-N (2r,3r,4s,5r,6s)-2-(hydroxymethyl)-6-[[(2r,3s,4r,5s,6r)-4,5,6-trihydroxy-3-[(2s,3s,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxan-2-yl]methoxy]oxane-3,4,5-triol 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]2[C@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)[C@H](O)[C@H](O)[C@H](O)O1 OMDQUFIYNPYJFM-XKDAHURESA-N 0.000 description 1
- XUCIJNAGGSZNQT-JHSLDZJXSA-N (R)-amygdalin 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[C@@H](C#N)C=2C=CC=CC=2)O1 XUCIJNAGGSZNQT-JHSLDZJXSA-N 0.000 description 1
- 229920001817 Agar Polymers 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
- 108091093088 Amplicon Proteins 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- HEBKCHPVOIAQTA-NGQZWQHPSA-N D-Arabitol Natural products OC[C@H](O)C(O)[C@H](O)CO HEBKCHPVOIAQTA-NGQZWQHPSA-N 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- HEBKCHPVOIAQTA-QWWZWVQMSA-N D-arabinitol Chemical compound OC[C@@H](O)C(O)[C@H](O)CO HEBKCHPVOIAQTA-QWWZWVQMSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 229920000926 Galactomannan Polymers 0.000 description 1
- 229920002907 Guar gum Polymers 0.000 description 1
- HEBKCHPVOIAQTA-IMJSIDKUSA-N L-arabinitol Chemical compound OC[C@H](O)C(O)[C@@H](O)CO HEBKCHPVOIAQTA-IMJSIDKUSA-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
- 229930195725 Mannitol Natural products 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
- 102100030569 Nuclear receptor corepressor 2 Human genes 0.000 description 1
- 101710153660 Nuclear receptor corepressor 2 Proteins 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- MUPFEKGTMRGPLJ-RMMQSMQOSA-N Raffinose Natural products O(C[C@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](O[C@@]2(CO)[C@H](O)[C@@H](O)[C@@H](CO)O2)O1)[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 MUPFEKGTMRGPLJ-RMMQSMQOSA-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
- MUPFEKGTMRGPLJ-UHFFFAOYSA-N UNPD196149 Natural products OC1C(O)C(CO)OC1(CO)OC1C(O)C(O)C(O)C(COC2C(C(O)C(O)C(CO)O2)O)O1 MUPFEKGTMRGPLJ-UHFFFAOYSA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 241001148470 aerobic bacillus Species 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 239000003513 alkali Substances 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
- 229940089837 amygdalin Drugs 0.000 description 1
- YZLOSXFCSIDECK-UHFFFAOYSA-N amygdalin Natural products OCC1OC(OCC2OC(O)C(O)C(O)C2O)C(O)C(O)C1OC(C#N)c3ccccc3 YZLOSXFCSIDECK-UHFFFAOYSA-N 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 229960000271 arbutin Drugs 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 238000005422 blasting Methods 0.000 description 1
- 230000005200 bud stage Effects 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000024346 drought recovery Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- YGHHWSRCTPQFFC-UHFFFAOYSA-N eucalyptosin A Natural products OC1C(O)C(O)C(CO)OC1OC1C(OC(C#N)C=2C=CC=CC=2)OC(CO)C(O)C1O YGHHWSRCTPQFFC-UHFFFAOYSA-N 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 239000000665 guar gum Substances 0.000 description 1
- 229960002154 guar gum Drugs 0.000 description 1
- 235000010417 guar gum Nutrition 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000015784 hyperosmotic salinity response Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 229920005610 lignin Polymers 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 239000010871 livestock manure Substances 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000005272 metallurgy Methods 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 238000005065 mining Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229950006780 n-acetylglucosamine Drugs 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- BJRNKVDFDLYUGJ-UHFFFAOYSA-N p-hydroxyphenyl beta-D-alloside Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=C(O)C=C1 BJRNKVDFDLYUGJ-UHFFFAOYSA-N 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000007639 printing Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 238000012113 quantitative test Methods 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
- 238000011084 recovery Methods 0.000 description 1
- 238000005067 remediation Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- CQRYARSYNCAZFO-UHFFFAOYSA-N salicyl alcohol Chemical compound OCC1=CC=CC=C1O CQRYARSYNCAZFO-UHFFFAOYSA-N 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 239000004753 textile Substances 0.000 description 1
Images
Classifications
-
- 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
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a method for ensiling sesbania and sweet sorghum in a mixing manner. The invention discloses a sesbania and sweet sorghum mixed ensiling method, which comprises the following steps: mixing the silage raw material with the compound lactobacillus preparation for solid anaerobic fermentation, and collecting a fermentation product to obtain sesbania silage; the ensiling raw material is a mixture consisting of sesbania aboveground parts and sweet sorghum aboveground parts; the active ingredients of the compound Lactobacillus preparation are Lactobacillus plantarum (Lactobacillus plantarum), lactobacillus hilgardii (Lactobacillus hilgardii) and Lactobacillus buchneri (Lactobacillus buchneri). The method can obviously prolong the aerobic stability of sesbania and sweet sorghum, fully decompose plant cell walls, utilize more sugar sources, maintain the pH and temperature of silage, keep the relative abundance of lactic acid bacteria in microbial flora and reduce the loss of dry matters.
Description
Technical Field
The invention relates to a method for ensiling mixed sesbania and sweet sorghum, belonging to the technical field of agricultural product processing.
Background
Sesbania (Sesbania cannabina) is a leguminous plant. Widely distributed in south areas such as China, hainan, jiangsu, zhejiang, jiangxi, fujian, guangxi, yunnan and the like, and north areas such as Shandong and the like are also cultivated. It is often found in wet and low-lying areas such as paddy fields and ditches. The sesbania has strong adaptability, salt tolerance, waterlogging tolerance, barren tolerance and drought tolerance, favors warmth and humidity, and is a pioneer crop for improving saline-alkali soil and soil remediation.
The sesbania is widely used, is an excellent green manure crop of various soil types, has seed endosperm capable of producing sesbania gum, is a novel galactomannan gum discovered in the 70 s of the 20 th century, and can replace imported guar gum. The sesbania gum has good water solubility, and is widely applied to industries such as oil extraction in oil fields, ore dressing metallurgy, mining blasting, textile printing and dyeing, medicines and pesticides, daily chemical industry and the like. The stem and leaf of sesbania can be used as livestock feed, but the research is less at present.
Generally, leguminous plants have high protein content and low soluble saccharide content, which cannot meet the basic requirements of ensiling, and how to ensile leguminous plants is a technical problem to be solved at present.
Disclosure of Invention
The technical problem to be solved by the method is to silage leguminous plants, especially sesbania.
In order to solve the above technical problems, the present invention first provides a method for preparing sesbania silage, comprising: mixing the silage raw material with the compound lactobacillus preparation to obtain a mixture before fermentation, performing solid anaerobic fermentation on the mixture before fermentation, and collecting a fermentation product to obtain sesbania silage;
the ensiling raw material is a mixture consisting of sesbania aboveground parts and sweet sorghum aboveground parts;
the active ingredients of the compound Lactobacillus preparation are Lactobacillus plantarum (Lactobacillus plantarum), lactobacillus hilgardii (Lactobacillus hilgardii) and Lactobacillus buchneri (Lactobacillus buchneri).
In the above method, the Lactobacillus plantarum (Lactobacillus plantarum) may be Lactobacillus plantarum (Lactobacillus plantarum) WQ-01;
the Lactobacillus hilgardii can be Lactobacillus hilgardii 60TS-2;
the Lactobacillus buchneri can be Lactobacillus buchneri (NX 205), and the Lactobacillus buchneri (NX 205) is a strain with the preservation number of CGMCC No.16534 in the China Committee for culture Collection of microorganisms.
In the composite Lactobacillus preparation of the above method, the ratio of the viable count of the Lactobacillus plantarum (Lactobacillus plantarum), the Lactobacillus hilgardii (Lactobacillus hilgardii) and the Lactobacillus buchneri (Lactobacillus buchneri) can be 1:1:1.
in one embodiment of the present invention, the complex Lactobacillus preparation is prepared from Lactobacillus plantarum (Lactobacillus plantarum), lactobacillus hilgardii (Lactobacillus hilgardii), lactobacillus buchneri (Lactobacillus buchneri) and physiological saline.
In the method, the mass percentage of the aboveground part of sesbania in the ensiling raw material can be 30-70%.
In the method, the aboveground part of the sesbania and the aboveground part of the sweet sorghum are both small sections of 2-4 cm.
The ratio of the ensiling raw material to the Lactobacillus plantarum (Lactobacillus plantarum), the Lactobacillus hilgardii (Lactobacillus hilgardii) and the Lactobacillus buchneri (Lactobacillus buchneri) in the composite Lactobacillus preparation in the pre-fermentation mixture in the above method may be 1:1:1.
in the above process, the fermentation may be carried out at a temperature of from 15 to 25 deg.C (e.g., from 20 to 25 deg.C). The fermentation time may be 14-60 days.
The fermentation can be carried out under vacuum, sealed and light-proof conditions, and particularly can be carried out in a sealed polyethylene bag.
The sesbania silage prepared by the method also belongs to the protection scope of the invention.
The compound lactobacillus preparation also belongs to the protection scope of the invention.
The invention also provides a composition for preparing sesbania silage, and the composition consists of the compound lactic acid bacteria preparation and the silage raw material.
Lactobacillus buchneri NX205 also belongs to the protection scope of the invention, and the preservation number of the Lactobacillus buchneri NX205 in the general microbiological center of China Committee for culture Collection of microorganisms is CGMCC No.16534.
The invention also provides a microbial inoculum, and the active ingredient of the microbial inoculum is the Lactobacillus buchneri (Lactobacillus buchneri) NX205.
The sesbania silage preparation method, or the sesbania silage, or the compound Lactobacillus preparation or the composition, or the Lactobacillus buchneri (Lactobacillus buchneri) NX205 or the application of the microbial inoculum in the preparation of animal feeds also belong to the protection scope of the invention.
The invention utilizes the compound Lactobacillus preparation (containing Lactobacillus plantarum WQ-01, lactobacillus hilgardii 60TS-2 and Lactobacillus buchneri NX 205) for mixed ensiling of sesbania and sweet sorghum to perform ensiling fermentation on the sesbania and the sweet sorghum, can obviously prolong the aerobic stability of the sesbania and the sweet sorghum, fully decompose plant cell walls, utilize more sugar sources, maintain the ensiling pH and temperature, maintain the relative abundance of Lactobacillus in microbial flora and reduce the dry matter loss.
Biological material preservation instructions
And (3) classification and naming: lactobacillus buchneri (Lactobacillus buchneri)
The strain number is as follows: NX205
Name of the depository: china general microbiological culture Collection center
The preservation unit is abbreviated as: CGMCC
The address of the depository: west road No.1 hospital No. 3, beijing, chaoyang district, zip code: 100101
The preservation date is as follows: 2018-9-26
Registration number of the preservation center: CGMCC No.16534
Drawings
FIG. 1 shows the effect of a composite silage bacterial agent on the pH value of a sesbania and sweet sorghum mixed silage sample.
FIG. 2 shows the effect of the composite silage bacterial agent on the lactic acid content of a sesbania and sweet sorghum mixed silage sample.
FIG. 3 is a graph showing the effect of a complex silage bacterial agent on the content of mixed silage acetic acid of sesbania and sweet sorghum samples.
FIG. 4 shows the effect of the composite silage bacterial agent on the content of 1, 2-propanediol in mixed silage of sesbania and sweet sorghum.
FIG. 5 shows the effect of the composite silage bacterial agent on the content of viable lactic acid bacteria in a mixed silage sample of sesbania and sweet sorghum.
FIG. 6 is a graph showing the effect of the composite silage bacterial agent on the culturable yeast content of a mixed silage sample of sesbania and sweet sorghum.
FIG. 7 shows the effect of the composite silage bacterial agent on the content of aerobic bacteria which can be cultured in a mixed silage sample of sesbania and sweet sorghum.
FIG. 8 shows the dynamic changes of microbial flora of horizontal sesbania and sweet sorghum mixed silage samples. D3, D7, D14, D30, D60 represent days 3, 7, 14, 30 and 60 of the silage fermentation, respectively.
Wherein SS100, SS70, SS50 and SS30 respectively represent sesbania proportions of 100%, 70%, 50% and 30%.
Detailed Description
The present invention is described in further detail below with reference to specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention. The examples provided below serve as a guide for further modifications by a person skilled in the art and do not constitute a limitation of the invention in any way.
The experimental procedures in the following examples, unless otherwise indicated, are conventional and are carried out according to the techniques or conditions described in the literature in the field or according to the instructions of the products. Materials, reagents, instruments and the like used in the following examples are commercially available unless otherwise specified. In the quantitative tests in the following examples, three replicates were set up and the results averaged.
Example 1 preparation of Mixed silage of Sesbania and sweet sorghum
The embodiment provides a compound Lactobacillus preparation for mixed silage of sesbania and sweet sorghum, which comprises Lactobacillus plantarum WQ-01 (Lactobacillus plantarum WQ-01 for short), lactobacillus hilgardii 60TS-2 (Lactobacillus hilgardii 60TS-2 for short) and Lactobacillus buchneri NX205 (Lactobacillus buchneri) for short, and the compound Lactobacillus preparation can remarkably prolong aerobic stability of the sesbania and the sweet sorghum, fully decompose plant cell walls, utilize more sugar sources, maintain silage pH and temperature, maintain relative abundance of Lactobacillus in microbial flora and reduce dry matter loss.
Wherein, the Lactobacillus plantarum WQ-01 and the Lactobacillus hilgardii 60TS-2 are both described in Chinese patent application No. 202010258231.8 (application publication No. CN 111534456A).
The lactobacillus buchneri NX205 is a strain NX205 separated from luocai group sweet sorghum in Ningxia Helan county, and the separation and identification steps are as follows:
1) Accurately weighing 10.00g of sweet sorghum sample by using a balance, fully mixing the sweet sorghum sample with 90mL of sterile water, shaking the mixture for 30min at 150rpm of a shaking table at room temperature, filtering the mixture by using four layers of gauze to obtain a leaching solution, uniformly coating the leaching solution on an MRS solid culture medium by using a sterilized coating rod after gradient dilution, and placing the MRS solid culture medium at 37 ℃ for anaerobic culture for 48h.
2) After the MRS solid culture medium grows out of the colonies, selecting white, smooth and convex colonies with neat edges by using a sterilized inoculating loop, inoculating the white, smooth and convex colonies into 3mL of MRS liquid culture medium, placing the MRS liquid culture medium at 37 ℃, and standing for anaerobic culture for 48 hours.
3) After the growth of bacteria in the MRS liquid medium, the bacteria liquid was transferred to a sterilized centrifuge tube by a 1mL pipette, centrifuged at 12000rpm for 3min, and then the bacteria was collected, and genomic DNA was extracted by Beijing Rui Boxing Ke Biotechnology GmbH (bacterial genomic DNA Rapid extraction kit, bomaide, cat #: DL 111-02) as a template, using a universal primer (27F: 5-: 5 '-TACGGCTACCTTGTTACAGACTT-3') 16S DNA was amplified by PCR and detected by 1% agarose gel electrophoresis. And (3) purifying and recovering the PCR amplification product by using a PCR product purification and recovery kit (Bomaide, product number: DH 102-02), and sequencing, wherein the sequence of the strain NX205 is a sequence 1 in a sequence table.
The strain NX205 is rod-shaped. The sequenced sequences are compared by using NCBI, the strain NX205 is identified as Lactobacillus buchneri (Lactobacillus buchneri), and is marked as Lactobacillus buchneri NX205, and the strain is preserved in China general microbiological culture Collection center (CGMCC) in 2018, 9 and 26 days, and the preservation number is CGMCC No.16534.
1. Characteristics of strain and sugar utilization characteristics
Lactobacillus plantarum WQ-01, lactobacillus hilgardii 60TS-2 and Lactobacillus buchneri NX205 were grown in MRS liquid medium and in MRS solid medium (1.5% agar powder added in MRS liquid medium), and anaerobically cultured at 37 ℃. The characteristics of the strain are shown in table 1, and the three are gram-positive bacteria, rod-shaped structures, catalase negative and oxidase negative. Lactobacillus plantarum WQ-01 is a homofermentative strain, capable of growing at ph 3.5-8.5, weaker at ph2.0, growing at 15-45 ℃, tolerant to 3.0% and 6.5% nacl concentration; lactobacillus hilgardii 60TS-2 is a heterotypic fermentation strain, growing at pH3.5-8.0, with weaker growth at pH2.0, at 15-40 deg.C, with weaker growth at 45 deg.C, tolerating 3.0% and 6.5% NaCl concentration; lactobacillus buchneri NX205 is a heterotypic fermentation strain that can grow at ph 4.0-4.5, weakly at ph2.0 and ph3.5, at 15-40 ℃, weakly at 45 ℃, tolerates 3.0% nacl concentration, and weakly at 6.5% nacl concentration.
The sugar utilization conditions are shown in table 2, and lactobacillus plantarum WQ-01 has the utilization capacity for ribose, galactose, glucose, fructose, mannose, mannitol, sorbitol, N-acetyl-glucosamine, amygdalin, arbutin, saligenin, cellobiose, maltose, lactose, melibiose, sucrose, trehalose, pinosyl, raffinose, taurochol, and L-arabitol, and grows weakly in a medium in which glycerol and D-arabitol are the only carbon sources, respectively; the lactobacillus hilgardii 60TS-2 has the utilization capacity on L-arabinose, ribose, D-xylose and sucrose, and has weak growth in a culture medium which respectively takes mannose, N-acetyl-glucose, cellobiose and sucrose as unique carbon sources; lactobacillus buchneri NX205 has the ability to use L-arabinose, ribose, D-xylose, glucose, maltose, melibiose, sucrose and pinosyl sugar, and has weak growth in media with galactose and fructose as the sole carbon sources, respectively. In Table 2, "control" means that the medium contained no carbon source.
TABLE 1 Lactobacillus plantarum WQ-01, lactobacillus hilgardii 60TS-2 and Lactobacillus buchneri NX205 Strain Properties
In Table 1, "W" indicates that there is weak growth.
TABLE 2 sugar utilization by Lactobacillus plantarum WQ-01, lactobacillus hilgardii 60TS-2 and Lactobacillus buchneri NX205
In Table 2, "W" indicates that there was weak growth.
2. Preparation of composite silage bacterial agent
Culturing the lactobacillus plantarum WQ-01 by using an MRS liquid culture medium (Beijing Soulebao science and technology Co., ltd., product number M8540-250 g), collecting a culture solution, centrifuging, collecting thalli, washing the thalli with physiological saline for three times, and collecting the thalli to obtain the lactobacillus plantarum WQ-01 microbial inoculum.
The lactobacillus hilgardii 60TS-2 is cultured by using MRS liquid culture medium (Beijing Soulebao science and technology Co., ltd., product number M8540-250 g), culture solution is collected, centrifugation is carried out, thalli are collected, and the thalli are washed by using physiological saline for three times and then are collected, thus obtaining the lactobacillus hilgardii 60TS-2 microbial inoculum.
The lactobacillus buchneri NX205 microbial inoculum is obtained by culturing lactobacillus buchneri NX205 by using an MRS liquid culture medium (Beijing Sorbao science and technology Co., ltd., product number M8540-250 g), collecting a culture solution, centrifuging, collecting the thalli, washing the thalli with physiological saline for three times, and collecting the thalli.
Mixing a lactobacillus plantarum WQ-01 bacterial agent, a lactobacillus hilgardii 60TS-2 bacterial agent and a lactobacillus buchneri NX205 bacterial agent according to a viable bacteria ratio of 1 8 cfu/g、10 8 cfu/g、10 8 cfu/g。
3. Preparation of sesbania and sweet sorghum mixed silage
Harvesting the fresh sesbania aboveground part and the sweet sorghum aboveground part in the bud stage, cutting the sesbania aboveground part and the sweet sorghum aboveground part respectively to a length of 2-4 cm (Kaiyue machinery company, model: 9 ZP-3.6).
Mixing and grouping the cut sesbania and sweet sorghum according to the table 3 respectively to obtain fresh plant samples; then adding a composite silage bacterial agent into each group of fresh plant samples to obtain a mixture before fermentation, wherein the viable bacteria content of the lactobacillus plantarum WQ-01, the lactobacillus hilgardii 60TS-2 and the lactobacillus buchneri NX205 in the mixture before fermentation is 10 8 CFU/g, the total mass of fresh plant samples is 300g.
Putting the mixture before fermentation into polyethylene bags, wherein the total mass of each bag of fresh plant sample is 300g, repeating for 4 groups, performing silage fermentation at room temperature (20-25 ℃) in a dark place after vacuumizing, sampling at 0 day, 3 day, 7 day, 14 day, 30 day and 60 day of the silage fermentation respectively, and detecting dry substances (DM) and silage fermentation products.
TABLE 3 grouping and blending ratio of Sesbania and sorghum saccharatum
4. Sample collection and detection
1. Dry Matter (DM) detection
Dry matter detection was performed by opening bags at 0, 3, 7, 14, 30 and 60 days of silage fermentation: weighing 100g of sample, drying the sample in a constant-temperature dryer at 65 ℃ to constant weight (72 h), naturally cooling the sample to room temperature, and weighing the sample, wherein the dry matter content (DM) = m/100 multiplied by 100 percent when the sample weight m is recorded; the samples were then assayed for Crude Protein (CP), neutral Detergent Fiber (NDF), acid Detergent Fiber (ADF), acid Detergent Lignin (ADL) and soluble carbohydrates (WSC). Basis of detection methodhttps://chinese.foragelab.com/Resources/Lab-ProceduresThe method is as follows.
2. Measurement of silage fermentation products
Opening bags at 0, 3, 7, 14, 30 and 60 days of silage fermentation for pH, lactic acid, acetic acid and 1, 2-propanediol assays: taking 10g of silage material, adding 90mL of normal saline, uniformly mixing, and placing in a refrigerator at 4 ℃ for standing overnight; the pH, lactic acid, acetic acid and 1, 2-propanediol were then measured in a 50mL centrifuge tube using a 4-layer gauze filter. The pH was measured using a pH meter (pH 213. Lactic acid, acetic acid and 1, 2-propanediol were detected by HPLC (1200, agilent, america) equipped with a UV detector (210 nm) and a column (ICSep COREGEL-87H) at 55 ℃ with a mobile phase of 0.005M H 2 SO 4 The flow rate of the aqueous solution was 0.6mL/min.
3. Culturable microbial flora detection
Opening bags at 0 day, 3 day, 7 day, 14 day, 30 day and 60 day of silage fermentation, adding 90mL of sterile PBS into 10g of silage samples, oscillating for 30min, filtering the solution with 4 layers of gauze, collecting filtrate (namely bacterial suspension) and placing the filtrate in a 50mL sterile centrifuge tube; and (3) diluting and coating the bacterial suspension, respectively coating the bacterial suspension in MRS, LB and PDA solid culture media, and counting microorganisms for about 36-48 h.
4. Microorganism analysis method based on high-throughput sequencing
Opening bags at 0, 3, 7, 14, 30 and 60 days of silage fermentation, taking 10.0g of sample for DNA extraction, carrying out SMRT sequencing, and determining the 16S rDNA full-length sequence. Microbial DNA was analyzed by paired-end sequencing (2 × 300 bp) using the Illumina MiSeq platform. The bacterial 16S rDNA amplicon sequencing used the V1-V9F (5. High quality sequences were obtained using QIIME quality control procedure and chimeric sequences were detected and deleted using UCHIME algorithm. And obtaining an OUT table, selecting each group of Species with the highest abundance ranking on each classification level (Phyum, class, order, family, genus, specifices) by adopting a maximum value ordering method according to Species annotation results, and generating a Species relative abundance column accumulation chart so as to visually check the microbial composition, relative abundance and proportion of each group on different classification levels.
5. Results and analysis
1. The influence of the composite silage bacterial agent on the silage chemical components of sesbania and sweet sorghum:
results as shown in table 4, the mixing ratio of sesbania and sorgo and the silage fermentation time together affect the DM, CP, NDF, ADF, ADL and WSC of the silage. The DM content of each combination silage remains relatively stable at each time point throughout the sesbania and sweet sorghum mixing silage process. The individual combined silage CP and silage ADL contents decrease significantly (P < 0.05) with increasing sweet sorghum addition, and each combined silage CP and silage ADL content decreases with fermentation time. The NDF content and the ADF content of each combined silage are not obviously related to the addition amount of the sweet sorghum, and the NDF content and the ADF content of each combined silage keep relatively stable at different time points. The WSC content of each combined silage is obviously increased along with the increase of the addition amount of the sweet sorghum (P is less than 0.05), and the WSC content of each combined silage is continuously reduced along with the fermentation time, so that the silage rule is also met.
TABLE 4 changes in DM, CP, NDF, ADF, ADL and WSC content during fermentation of sesbania and sorghum saccharatum mixed silage
In Table 4, different lower case letters indicate significant differences in nutritional quality (p < 0.05) for different samples at the same fermentation time, and different upper case letters indicate significant differences in nutritional quality (p < 0.01) for the same sample at different fermentation times.
2. The influence of the composite silage bacterial agent on silage fermentation products of sesbania and sweet sorghum:
as shown in fig. 1 and table 5, compared with the SS100 combination, the pH values of the SS70 combination, the SS50 combination and the SS30 combination all have obvious falling tendency within 60 days of ensiling fermentation, which indicates that even if lactic acid bacteria are added, the pH value of sesbania ensiling fermentation alone is difficult to reduce, and the addition of sweet sorghum in sesbania can well exert the advantages of lactic acid bacteria and effectively maintain the pH of sesbania and sweet sorghum mixed ensiling.
As shown in fig. 2 and table 5, the lactic acid content of the SS30 combination, the SS50 combination, and the SS70 combination was higher than that of the SS100 combination at each period of time, and the lactic acid content of the SS30 combination was the highest as a whole, followed by the SS50 combination, within 60 days of the silage fermentation.
As shown in fig. 3 and table 5, the SS100 group silage (100% sesbania group) had the highest acetic acid content at four time points within 30 days of fermentation, followed by the SS30 combination. Within 14 days of silage fermentation, the acetic acid content of the SS70, SS50 and SS30 combined silage is slightly changed and kept stable. After 14 days of silage fermentation, the acetic acid content of the SS100 combined silage is gradually reduced, and the acetic acid content of the SS70, SS50 and SS30 combined silage is rapidly increased.
As shown in fig. 4 and table 5, 1, 2-propanediol was detected at day 14 of fermentation, but as fermentation time increased, the 1, 2-propanediol content of the SS100 combination silage consistently increased faster than the 1, 2-propanediol content of the 0, ss70, SS50 and SS30 combination silage, and the 1, 2-propanediol content of the SS30 combination silage was highest with a high proportion of sweet sorghum added.
TABLE 5 influence of the Compound silage inoculant on the ensiling fermentation products of Sesbania and sweet sorghum for 60 days of ensiling fermentation
Different lower case letters indicate significant differences in fermentation quality (p < 0.05) for different samples at the same fermentation time, and different upper case letters indicate significant differences in fermentation quality (p < 0.01) for the same sample at different fermentation times.
3. The influence of the composite silage bacterial agent on the cultivated microbial flora of the silage and sweet sorghum silage is as follows:
as shown in fig. 5, the contents of the four combinations of culturable lactic acid bacteria were approximately the same in the variation trend within 60 days of ensiling, wherein the content of the SS100 combination of culturable lactic acid bacteria was always the highest at each time point.
As shown in fig. 6, the culturable yeast content of the four combinations varied approximately equally over 60 days of ensiling, with the culturable yeast content of the SS100 combination being the highest at each time point.
As shown in FIG. 7, the culturable aerobic bacterial contents of the four combinations varied approximately in the same manner during the 60 days of ensiling, with the culturable yeast content of the SS100 combination being the highest at each time point.
4. The influence of the composite silage bacterial agent on the high-throughput sequencing floras of the silage microorganisms of sesbania and sweet sorghum:
species relative abundance bar graphs (species levels) for sesbania and sweet sorghum mixed silage bacterial communities are shown in fig. 8, tables 6-8. Before ensiling, sesbania fresh-like and sweet sorghum fresh-like floras are relatively complex, the relative abundance of Bacillus subtilis in the sesbania fresh-like is 41.29 percent, the most dominant strain is, and the relative abundance of the Bacillus subtilis is Stenotrophormonas maltophia with 6.06 percent. The relative abundance of Bacillus subtilis in the fresh sweet sorghum sample is 31.83 percent, and the strain is the most dominant strain. After ensiling, each strain in the composite ensiling microbial inoculum exerts the dominant advantages, but the action time is different. In the early stage of fermentation (before day 14), homotype lactobacillus plantarum has dominant advantages, and in the later stage of fermentation (after day 14), heterotype lactobacillus hilgardii and lactobacillus buchneri have dominant advantages after replacing homotype lactobacillus, and the trend of the above rules is more obvious in SS70, SS50 and SS30 combined silage. Whereas the trend of change was less pronounced in SS100 combination silage. After the strain is ensiled and fermented in SS100 combined silage for 60 days, the relative abundances of the three lactobacillus added strains, namely lactobacillus hilgardii, lactobacillus buchneri and lactobacillus plantarum are respectively 10.63%, 7.29% and 11.54%. While the relative abundance of lactobacillus hilgardii in the combined SS70, SS50 and SS30 silage was 33.13%, 28.75% and 67.58%, and the relative abundance of lactobacillus buchneri in the combined SS70, SS50 and SS30 silage was 62.46%, 65.93% and 29.47%, respectively.
TABLE 6 relative abundance of microbial flora before and after mixed ensiling of horizontal sesbania and sweet sorghum
TABLE 7 relative abundance of microbial flora before and after mixed ensiling of sesbania and sweet sorghum at levels
TABLE 8 relative abundance of microbial flora before and after mixed ensiling of horizontal sesbania and sorgo
The present invention has been described in detail above. It will be apparent to those skilled in the art that the invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with reference to specific embodiments, it will be appreciated that the invention can be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains. The use of some of the essential features is possible within the scope of the claims attached below.
<110> institute for microbiology of Chinese academy of sciences
<120> a method for ensiling sesbania and sweet sorghum mixture
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 1521
<212> DNA
<213> Lactobacillus buchneri (Lactobacillus buchneri)
<220>
<221> misc_feature
<222> (1)..(2)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (136)..(136)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (154)..(154)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (197)..(197)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (225)..(225)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (231)..(233)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (272)..(272)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (371)..(373)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (387)..(387)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (562)..(563)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (582)..(582)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (648)..(648)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (772)..(773)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (777)..(777)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (779)..(779)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (969)..(969)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (1003)..(1004)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (1026)..(1026)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (1424)..(1425)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (1438)..(1440)
<223> n is a, c, g, or t
<220>
<221> misc_feature
<222> (1485)..(1485)
<223> n is a, c, g, or t
<400> 1
nntaagatga gagtttgatc ctggctcagg acgaacgctg gcggcgtgcc taatacatgc 60
aagtcgaacg cgtctccgtt ratgayttta rgtgcttgca yttgaaagat ttaacattga 120
gacgagtggc gaactngtga gtaacacgtg ggtnacctgc ccttgaagta ggggataaca 180
cttggaaaca ggtgctnata ccgtataaca accaaaacct acctngtttt nnnttaaaag 240
acggcttcgg ctgtcacttt aggatggacc cncggcgtat tagcttgttg gtaaggtaac 300
ggcctaccaa ggcgatgata cgtagccgac ctgagagggt aatcggccac attgggactg 360
agacacggcc nnnactccta cgggagncag cagtagggaa tcttccacaa tggacgaaag 420
tctgatggag caacgccgcg tgagtgatga agggtttcgg ctcgtaaaac tctgttgttg 480
gagaagaaca ggtgtcagag yaactgttga catcttgacg gtatccaacc agaaagccac 540
ggctaactac gtgccagcag cnncggtaat acgtaggtgg cnagcgttgt ccggatttat 600
tgggcgtaaa gcgagcgcag gcggtttttt aggtctgatg tgaaagcntt cggcttaacc 660
ggagaagtgc atcggaaacc gggaggcttg agtgcagaag aggacagtgg aactccatgt 720
gtagcggtga aatgcgtaga tatatggaag aacaccagtg gcgaaggcgg cnntctngnc 780
tgtaactgac gctgaggctc gaaagcatgg gtagcgaaca ggattagata ccctggtagt 840
ccatgccgta aacgatgagt gctaagtgtt ggagggtttc cgcccttcag tgctgcagct 900
aacgcattaa gcactccgcc tggggagtac gaccgcaagg ttgaaactca aaggaattga 960
cgggggccng cacaagcggt ggagcatgtg gtttaattcg atnntacgcg aagaacctta 1020
ccaggncttg acatcttctg ccaacctaag agattaggcg ttcccttcgg ggacagaatg 1080
acaggtggtg catggttgtc gtcagctcgt gtcgtgagat gttgggttaa gtcccgcaac 1140
gagcgcaacc cttattgtta gttgccagca ttcagttggg cactctagca agactgccgg 1200
tgacaaaccg gaggaaggtg gggatgacgt caaatcatca tgccccttat gacctgggct 1260
acacacgtgc tacaatggac ggtacaacga gtcgcgaaac cgcgaggtca agctaatctc 1320
ttaaagccgt tctcagttcg gattgtaggc tgcaactcgc ctacatgaag ttggaatcgc 1380
tagtaatcgt ggatcagcat gccacggtga atacgttccc gggnnttgta cacaccgnnn 1440
gtcacaccat gagagtttgt aacacccaaa gccggtgagg taacnttcgg ggaccagccg 1500
tctaaggtgg gacagatgat t 1521
Claims (10)
1. A method of preparing sesbania silage, comprising: mixing the silage raw material with the compound lactobacillus preparation to obtain a mixture before fermentation, performing solid anaerobic fermentation on the mixture before fermentation, and collecting a fermentation product to obtain sesbania silage;
the ensiling raw material is a mixture consisting of sesbania aboveground parts and sweet sorghum aboveground parts;
the active ingredients of the compound Lactobacillus preparation are Lactobacillus plantarum (Lactobacillus plantarum), lactobacillus hilgardii (Lactobacillus hilgardii) and Lactobacillus buchneri (Lactobacillus buchneri).
2. The method of claim 1, wherein: the Lactobacillus plantarum (Lactobacillus plantarum) is Lactobacillus plantarum WQ-01;
the Lactobacillus hilgardii is Lactobacillus hilgardii 60TS-2;
the Lactobacillus buchneri is Lactobacillus buchneri (Lactobacillus buchneri) NX205, and the Lactobacillus buchneri (Lactobacillus buchneri) NX205 is a strain with the preservation number of CGMCC No.16534 in the China general microbiological culture Collection center.
3. The method according to claim 1 or 2, characterized in that: in the composite Lactobacillus preparation, the ratio of the viable count of the Lactobacillus plantarum (Lactobacillus plantarum), the Lactobacillus hilgardii (Lactobacillus hilgardii) and the Lactobacillus buchneri (Lactobacillus buchneri) is 1:1:1.
4. a method according to any one of claims 1-3, characterized in that: the mass percentage of the overground part of the sesbania in the ensiling raw material is 30-70%.
5. The method according to any one of claims 1-4, wherein: in the pre-fermentation mixture, the ratio of the ensiling raw material to the Lactobacillus plantarum (Lactobacillus plantarum), the Lactobacillus hilgardii (Lactobacillus hilgardii) and the Lactobacillus buchneri (Lactobacillus buchneri) in the composite Lactobacillus preparation is 1:1:1.
6. the method according to any one of claims 1 to 5, wherein: the fermentation is carried out at 15-25 ℃;
and/or the fermentation time is 14-60 days.
7. Sesbania silage produced by the method of any one of claims 1-6.
8. The complex lactic acid bacteria preparation of any one of claims 1 to 3;
or, a composition for preparing sesbania silage, consisting of the composite lactic acid bacteria preparation of any one of claims 1 to 3 and the silage material of any one of claims 1 to 4.
9. Lactobacillus buchneri (Lactobacillus buchneri) NX205 with the preservation number of CGMCC No.16534 in China general microbiological culture Collection center;
or a microbial inoculum, the active ingredient of which is the Lactobacillus buchneri (Lactobacillus buchneri) NX205.
10. Use of the method of any one of claims 1 to 6, or the sesbania ensilage of claim 7, or the complex lactic acid bacteria preparation of claim 8, or the composition, or the Lactobacillus buchneri (Lactobacillus buchneri) NX205 of claim 9, or the microbial inoculum for the preparation of animal feed.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110907045.7A CN115702667B (en) | 2021-08-09 | 2021-08-09 | Mixed silage method for sesbania and sweet sorghum |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110907045.7A CN115702667B (en) | 2021-08-09 | 2021-08-09 | Mixed silage method for sesbania and sweet sorghum |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115702667A true CN115702667A (en) | 2023-02-17 |
| CN115702667B CN115702667B (en) | 2024-03-08 |
Family
ID=85179403
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110907045.7A Active CN115702667B (en) | 2021-08-09 | 2021-08-09 | Mixed silage method for sesbania and sweet sorghum |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115702667B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108935972A (en) * | 2018-05-14 | 2018-12-07 | 中国科学院微生物研究所 | A kind of method of feeding soybean and the mixed storage of sugar grass |
-
2021
- 2021-08-09 CN CN202110907045.7A patent/CN115702667B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108935972A (en) * | 2018-05-14 | 2018-12-07 | 中国科学院微生物研究所 | A kind of method of feeding soybean and the mixed storage of sugar grass |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115702667B (en) | 2024-03-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111876351B (en) | Bacillus belgii and application thereof in relieving apple continuous cropping obstacle | |
| CN107312734B (en) | Banana wilt antagonistic bacteria and bio-organic fertilizer as well as preparation method and application thereof | |
| Scheuerell et al. | Compost tea as a container medium drench for suppressing seedling damping-off caused by Pythium ultimum | |
| CN106754557B (en) | Bacillus subtilis YBM-4 and application thereof in preventing and treating tobacco black shank and promoting growth | |
| CN110591988B (en) | Lactobacillus rhamnosus753 and application thereof, silage additive and silage | |
| CN108893421B (en) | Bacillus fusiformis and application thereof in reclamation ecological reconstruction of mining area | |
| CN112080435A (en) | Purple lilac spore strain and application thereof in degrading chicken feather | |
| CN109182181B (en) | High-temperature micro-aerobic composting nitrogen-retaining bacterium for reducing ammonia volatilization of livestock and poultry manure compost and application thereof | |
| CN108060102B (en) | A kind of cibarium Wei Si Salmonella and its application, screening calibration method | |
| CN106635926B (en) | Bacillus safensis YJC-4 and application thereof in prevention and treatment of tobacco black shank and growth promotion | |
| Thompson et al. | Survival of two ecologically distinct bacteria (Flavobacterium and Arthrobacter) in unplanted and rhizosphere soil: laboratory studies | |
| CN112877238A (en) | Low-temperature-resistant pediococcus pentosaceus OL77 and application thereof | |
| CN114908017B (en) | Lactobacillus plantarum and application thereof in silage | |
| CN115702667B (en) | Mixed silage method for sesbania and sweet sorghum | |
| CN113604399B (en) | Sphingolipid bacteria with growth promoting function of garden plants and application thereof | |
| CN112795520B (en) | Bacillus for degrading manure | |
| CN112715758B (en) | Lactobacillus buchneri BL4 and method for preparing silage by using same | |
| CN115702668A (en) | Method for mixed ensiling of cyperus esculentus stem leaves and rape straws | |
| CN111040969B (en) | Compound lactobacillus agent and application thereof in buffalo silage | |
| CN114806924B (en) | Achromobacter denitrificans and application thereof | |
| Fujii et al. | Combined use of cyanoditolyl tetrazolium staining and flow cytometry for detection of metabolically active bacteria in a fed-batch composting process | |
| CN114634889B (en) | Bacillus licheniformis nmxc2-1, fermentation product, microbial inoculum and application thereof | |
| CN117165478B (en) | Preparation method and application of bacillus belicus SL-K2, biocontrol agent and gingerol extract | |
| CN110669705B (en) | Lysinibacillus fusiformis CA1 and application thereof in promoting growth of pleurotus geesteranus | |
| CN112300958B (en) | Hydroxyl bacteria S-1-4 and screening method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |