CN111304112B - Composite microbial inoculum for ensiling sugarcane tail leaves - Google Patents
Composite microbial inoculum for ensiling sugarcane tail leaves Download PDFInfo
- Publication number
- CN111304112B CN111304112B CN201911305751.3A CN201911305751A CN111304112B CN 111304112 B CN111304112 B CN 111304112B CN 201911305751 A CN201911305751 A CN 201911305751A CN 111304112 B CN111304112 B CN 111304112B
- Authority
- CN
- China
- Prior art keywords
- lactobacillus
- casei
- candida
- paracasei
- lactobacillus plantarum
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 240000000111 Saccharum officinarum Species 0.000 title claims abstract description 55
- 235000007201 Saccharum officinarum Nutrition 0.000 title claims abstract description 55
- 239000002131 composite material Substances 0.000 title claims abstract description 18
- 239000002068 microbial inoculum Substances 0.000 title claims abstract description 18
- 244000199866 Lactobacillus casei Species 0.000 claims abstract description 54
- 235000013958 Lactobacillus casei Nutrition 0.000 claims abstract description 44
- 241000186605 Lactobacillus paracasei Species 0.000 claims abstract description 44
- 229940017800 lactobacillus casei Drugs 0.000 claims abstract description 44
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 41
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 41
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 41
- 230000001580 bacterial effect Effects 0.000 claims abstract description 25
- 241000222173 Candida parapsilosis Species 0.000 claims abstract description 23
- 229940055022 candida parapsilosis Drugs 0.000 claims abstract description 23
- 241000222120 Candida <Saccharomycetales> Species 0.000 claims abstract description 19
- 239000004460 silage Substances 0.000 claims description 29
- 241000894006 Bacteria Species 0.000 claims description 27
- 239000003223 protective agent Substances 0.000 claims description 27
- 239000001963 growth medium Substances 0.000 claims description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 14
- 238000000034 method Methods 0.000 claims description 14
- 238000012258 culturing Methods 0.000 claims description 13
- 238000009630 liquid culture Methods 0.000 claims description 13
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 12
- 239000007788 liquid Substances 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- 238000002156 mixing Methods 0.000 claims description 10
- 239000000203 mixture Substances 0.000 claims description 9
- 238000002360 preparation method Methods 0.000 claims description 9
- 238000004108 freeze drying Methods 0.000 claims description 8
- 241000186660 Lactobacillus Species 0.000 claims description 7
- 229940039696 lactobacillus Drugs 0.000 claims description 7
- 230000003213 activating effect Effects 0.000 claims description 6
- 238000000855 fermentation Methods 0.000 claims description 6
- 230000004151 fermentation Effects 0.000 claims description 6
- 230000001954 sterilising effect Effects 0.000 claims description 6
- 239000006228 supernatant Substances 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 5
- 238000004659 sterilization and disinfection Methods 0.000 claims description 5
- 239000000725 suspension Substances 0.000 claims description 5
- 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 claims description 4
- 229920001817 Agar Polymers 0.000 claims description 4
- 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 claims description 4
- 239000008272 agar Substances 0.000 claims description 4
- 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 claims description 4
- 239000011248 coating agent Substances 0.000 claims description 4
- 238000000576 coating method Methods 0.000 claims description 4
- 239000013078 crystal Substances 0.000 claims description 4
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 4
- 239000002609 medium Substances 0.000 claims description 4
- 235000013923 monosodium glutamate Nutrition 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 235000020183 skimmed milk Nutrition 0.000 claims description 4
- 229940073490 sodium glutamate Drugs 0.000 claims description 4
- 238000007710 freezing Methods 0.000 claims description 3
- 230000008014 freezing Effects 0.000 claims description 3
- 230000000813 microbial effect Effects 0.000 claims description 3
- 238000004806 packaging method and process Methods 0.000 claims description 3
- 239000003755 preservative agent Substances 0.000 claims description 3
- 230000002335 preservative effect Effects 0.000 claims description 3
- 238000005070 sampling Methods 0.000 claims description 3
- 238000007789 sealing Methods 0.000 claims description 3
- 239000008223 sterile water Substances 0.000 claims description 3
- 239000002054 inoculum Substances 0.000 claims description 2
- 238000011068 loading method Methods 0.000 claims description 2
- 238000005507 spraying Methods 0.000 claims description 2
- 230000008021 deposition Effects 0.000 claims 4
- 230000003068 static effect Effects 0.000 claims 1
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 abstract description 18
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 abstract description 9
- 239000004310 lactic acid Substances 0.000 abstract description 9
- 235000014655 lactic acid Nutrition 0.000 abstract description 9
- 238000009395 breeding Methods 0.000 abstract description 2
- 230000001488 breeding effect Effects 0.000 abstract description 2
- 239000004480 active ingredient Substances 0.000 abstract 1
- 244000005700 microbiome Species 0.000 abstract 1
- 238000011156 evaluation Methods 0.000 description 8
- 238000009472 formulation Methods 0.000 description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- 239000003795 chemical substances by application Substances 0.000 description 6
- 230000001953 sensory effect Effects 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 5
- 239000000706 filtrate Substances 0.000 description 5
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 4
- 230000004913 activation Effects 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 238000013441 quality evaluation Methods 0.000 description 4
- 239000004459 forage Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000007747 plating Methods 0.000 description 3
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- 102000016938 Catalase Human genes 0.000 description 2
- 108010053835 Catalase Proteins 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-M Lactate Chemical compound CC(O)C([O-])=O JVTAAEKCZFNVCJ-UHFFFAOYSA-M 0.000 description 2
- LCTONWCANYUPML-UHFFFAOYSA-N Pyruvic acid Chemical compound CC(=O)C(O)=O LCTONWCANYUPML-UHFFFAOYSA-N 0.000 description 2
- 241000282849 Ruminantia Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 235000010855 food raising agent Nutrition 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- 235000013336 milk Nutrition 0.000 description 2
- 239000008267 milk Substances 0.000 description 2
- 210000004080 milk Anatomy 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000007873 sieving Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 1
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical class CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 1
- 241000222122 Candida albicans Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 238000003794 Gram staining Methods 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 240000001929 Lactobacillus brevis Species 0.000 description 1
- 235000013957 Lactobacillus brevis Nutrition 0.000 description 1
- UEZVMMHDMIWARA-UHFFFAOYSA-N Metaphosphoric acid Chemical compound OP(=O)=O UEZVMMHDMIWARA-UHFFFAOYSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 238000013494 PH determination Methods 0.000 description 1
- 235000015696 Portulacaria afra Nutrition 0.000 description 1
- 244000177175 Typha elephantina Species 0.000 description 1
- 235000018747 Typha elephantina Nutrition 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- VTYYLEPIZMXCLO-UHFFFAOYSA-L calcium carbonate Substances [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 229940095731 candida albicans Drugs 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 239000013256 coordination polymer Substances 0.000 description 1
- LDHQCZJRKDOVOX-NSCUHMNNSA-N crotonic acid Chemical compound C\C=C\C(O)=O LDHQCZJRKDOVOX-NSCUHMNNSA-N 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 238000006356 dehydrogenation reaction Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 235000013325 dietary fiber Nutrition 0.000 description 1
- 238000007598 dipping method Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000000556 factor analysis Methods 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 150000002431 hydrogen Chemical class 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000004311 natamycin Substances 0.000 description 1
- 235000010298 natamycin Nutrition 0.000 description 1
- NCXMLFZGDNKEPB-FFPOYIOWSA-N natamycin Chemical compound O[C@H]1[C@@H](N)[C@H](O)[C@@H](C)O[C@H]1O[C@H]1/C=C/C=C/C=C/C=C/C[C@@H](C)OC(=O)/C=C/[C@H]2O[C@@H]2C[C@H](O)C[C@](O)(C[C@H](O)[C@H]2C(O)=O)O[C@H]2C1 NCXMLFZGDNKEPB-FFPOYIOWSA-N 0.000 description 1
- 229960003255 natamycin Drugs 0.000 description 1
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 1
- BOPGDPNILDQYTO-NNYOXOHSSA-N nicotinamide-adenine dinucleotide Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 BOPGDPNILDQYTO-NNYOXOHSSA-N 0.000 description 1
- 235000021049 nutrient content Nutrition 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 229940107700 pyruvic acid Drugs 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000010802 sludge Substances 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- LDHQCZJRKDOVOX-UHFFFAOYSA-N trans-crotonic acid Natural products CC=CC(O)=O LDHQCZJRKDOVOX-UHFFFAOYSA-N 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K30/00—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs
- A23K30/10—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder
- A23K30/15—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder using chemicals or microorganisms for ensilaging
- A23K30/18—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder using chemicals or microorganisms for ensilaging using microorganisms or enzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/125—Casei
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/165—Paracasei
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Mycology (AREA)
- Tropical Medicine & Parasitology (AREA)
- General Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Animal Husbandry (AREA)
- Botany (AREA)
- Food Science & Technology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to the field of microorganisms, and particularly relates to a composite microbial inoculum for ensiling sugarcane tail leaves, which is characterized by comprising the following components in parts by weight: its active ingredients include Candida parapsilosis (Candida humils), Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus casei (Lactobacillus casei), and Lactobacillus paracasei (Lactobacillus paracasei). The inventor adopts the form of 4 dominant bacterial strains obtained by independent breeding of the applicant to combine the composite microbial inoculum, and after the fresh sugarcane tail leaves are subjected to ensiling treatment, compared with the form of using any single bacterial strain as the microbial inoculum, the quality of ensiling of the fresh sugarcane tail leaves is further improved, such as remarkably improving the content of lactic acid and acetic acid, remarkably reducing the pH value and reducing NH3-N content.
Description
Technical Field
The invention belongs to the field of biology, and particularly relates to a composite microbial inoculum for ensiling sugarcane tail leaves.
Background
In recent years, the demand for milk and meat of ruminants has been rapidly increased in many subtropical and tropical countries and regions, and the ruminant industry has also been rapidly developed, thereby causing a huge demand for roughage resources, and problems of shortage and unbalanced supply of high-quality forage are increasingly prominent.
Sugarcane is a gramineous plant widely grown in tropical and subtropical regions. The total national sugarcane yield in 2017 reaches 10440.4 ten thousand tons, wherein the total Guangxi yield reaches 7132.3 ten thousand tons and accounts for about 68.31 percent of the total national yield. The sugarcane tip is a general name of 2-3 tender knots and green leaves on the top of the sugarcane when the sugarcane is harvested, is an accessory product generated in the sugarcane planting process, and accounts for about 10% of the biomass of the sugarcane. The annual sugarcane tips produced in Guangxi areas are about 1400 million tons and huge in quantity, but less than 30 percent of sugarcane tips are used as feed to directly feed herbivorous animals, and most of sugarcane tips are burnt and discarded, so that the problems of great resource waste and serious environmental pollution are caused. The harvest season of the sugarcane is usually from 11 months to 5 months of the next year, and the forage application of the sugarcane tips can just make up the problem of the forage resource shortage caused by slow growth of the pasture in autumn and winter.
At present, the sugarcane tips are mainly directly and freshly fed or ensiled to feed in China, and more sugarcane tail ensiling methods are published. But the main method is to use lactic acid milk as a microbial additive. The invention discloses a method for preparing leaven with different addition proportions by using three dominant lactic acid bacteria and one yeast extracted from sugarcane tails to obtain better silage quality.
Disclosure of Invention
The invention aims to provide a compound microbial inoculum for ensiling sugarcane tail leaves. According to the invention, anaerobic ensiling is carried out by Candida parapsilosis (Candida humils), Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus casei (Lactobacillus casei) and Lactobacillus paracasei (Lactobacillus paracasei) which are independently developed by the research laboratory, so that the fermentation speed can be effectively promoted, the pH value of ensiling feed can be reduced, and the quantity of lactic acid and organic acid can be increased, thereby improving the ensiling quality of sugarcane tail leaves.
The invention content of the invention is as follows:
a composite bacterial preparation for ensiling sugarcane tail leaves comprises active components of Candida parapsilosis (Candida humils), Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus casei (Lactobacillus casei) and Lactobacillus paracasei (Lactobacillus paracasei).
The Candida parapsilosis (Candida humils) GS2 is preserved in China, Wuhan university, China center for type culture Collection in 7 months and 02 days in 2018, and the preservation number is CCTCC M2018441; lactobacillus plantarum GS3 was deposited in China, Wuhan university, China center for type culture Collection in 2018, 08-03 month, with a collection number of CCTCC M2018522; lactobacillus casei (Lactobacillus casei) GS4 was deposited in China, Wuhan university, China center for type culture Collection in 2018, 08-03 month, with a collection number of CCTCC M2018523; lactobacillus paracasei (Lactobacillus paracasei) GS5 was deposited in China, Wuhan university, China center for type culture Collection in 2018, 08-03.month with the collection number of CCTCC M2018524.
In the invention, the ratio of the Candida parapsilosis (Candida humils), the Lactobacillus plantarum (Lactobacillus plantarum), the Lactobacillus casei (Lactobacillus casei) and the Lactobacillus paracasei (Lactobacillus paracasei) is 0-100 percent, 0-10 percent and 0-10 percent in terms of viable count.
Preferably, the ratio of the Candida parapsilosis (Candida humils), the Lactobacillus plantarum (Lactobacillus plantarum), the Lactobacillus casei (Lactobacillus casei) and the Lactobacillus paracasei (Lactobacillus paracasei) is 20% -50%: 30% -60%: 10%:10% in terms of viable count percentage.
The preparation method of the composite microbial inoculum for ensiling the sugarcane tail leaves comprises the following steps:
(1) and activating: respectively inoculating the extracted 4 strains into liquid culture medium according to 10% of the addition amount of the strains, culturing Candida parapsilosis (Candida humils) at 28 deg.C for 24h, and performing shake culture at 150 r/min; the temperature of Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus casei (Lactobacillus casei) and Lactobacillus paracasei (Lactobacillus paracasei) is 37 ℃ for 24 hours, and the Lactobacillus plantarum, the Lactobacillus plantarum and the Lactobacillus paracasei are statically cultured;
(2) and (3) expanding culture: then respectively inoculating the activated 4 strains into a liquid culture medium according to the addition of 5 percent, and carrying out shake culture at 150r/min at the culture temperature of 28 ℃ for 24h by using Candida parapsilosis; culturing Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus casei (Lactobacillus casei) and Lactobacillus paracasei (Lactobacillus paracasei) at 37 ℃ for 24h, statically culturing, and counting inoculated bacteria liquid by coating;
(3) and (3) freeze drying: wiping the inner wall of a centrifuge with 95% ethanol to perform sterilization treatment, mixing concentrated and cultured bacteria fermentation liquor according to the concentration proportion of 4 strains by using a sterilization centrifugal tube, centrifuging for 20min at 3000r/min, discarding supernatant to obtain bacteria mud, fully mixing the bacteria mud with a proper amount of sterile water to prepare bacteria suspension with uniform concentration, and adding a protective agent, namely the bacteria mud: and (3) transferring the protective agents into a sterile culture dish together with the protective agents with the thickness of 1:3, wrapping the protective agents with a preservative film, pre-freezing the protective agents in an ultralow temperature refrigerator for 5 hours to form small ice crystals, immediately transferring the protective agents into a freeze drying device for drying until no clear water exists, sampling to determine the viable count, immediately carrying out vacuum sealing packaging, and then preserving the protective agents with a 4-degree refrigerator.
In the present invention, further explained are: the protective agent comprises, by percentage, 80% of fat-free milk powder, 8% of trehalose, 8% of sodium glutamate and 4% of glycerol.
In the present invention, further explained are: in the steps (1) to (2), the MRS liquid medium is used for activating Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus casei (Lactobacillus casei) and Lactobacillus paracasei (Lactobacillus paracasei) and culturing bacterial liquid; MRS solid culture medium is used for coating and counting Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus casei (Lactobacillus casei) and Lactobacillus paracasei (Lactobacillus paracasei); the YPD liquid culture medium is used for activating Candida parapsilosis (Candida humils) and culturing bacterial liquid; YPD agar was used for plating and counting Candida parapsilosis (Candida humils).
The application of the composite microbial inoculum for silage of sugarcane tail leaves in preparation of silage sugarcane tail leaves.
In the invention, the use method of the composite bacteria for ensiling the sugarcane tail leaves further comprises the following steps: cutting fresh sugarcane tail leaves into 2-3cm, spraying the composite microbial inoculum for ensiling the sugarcane tail leaves on the fresh sugarcane tail leaves, uniformly mixing, loading into an ensiling bag, vacuumizing, and fermenting for 60 days.
Compared with the prior art, the beneficial effect of this application:
the applicant finds that the quality of the silage of the fresh sugarcane tail leaves is further improved by adopting a form of 4 dominant bacterial strains obtained by independent breeding of the applicant and combining the composite bacterial agents compared with a form of any single bacterial strain serving as the bacterial agent after the silage treatment of the fresh sugarcane tail leaves, such as the contents of lactic acid and acetic acid are obviously improved, the pH value is obviously reduced, and NH is reduced3-N content.
Detailed Description
In order to make the aforementioned objects, features and advantages of the present invention more comprehensible, specific embodiments thereof will be described in detail with reference to the following examples. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein, but rather should be construed as broadly as the present invention is capable of modification in various respects, all without departing from the spirit and scope of the present invention.
Example 1
Candida parapsilosis (Candida humils), Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus casei (Lactobacillus casei), Lactobacillus paracasei (Lactobacillus paracasei) strains were obtained and named:
picking out the silage sugarcane tails by an inoculating loop, and adding 0.5 percent of CaCO3And lactobacillus plantarum colonies growing on an MRS solid medium plate added with 0.001% (g/100ml) of natamycin, randomly selecting colonies with smooth surfaces, round, milky white or grey white shapes and obvious caldolytic rings, and then respectively carrying out catalase detection (taking a piece of glass slide cleaned in advance, dripping a drop of 3% hydrogen peroxide, dipping the colonies in hydrogen peroxide liquid by using an inoculating loop, observing whether bubbles are generated, and whether the bubbles are generated to be positive or negative or not) and gram staining microscopy; selecting catalase as negative, gramBacterial colonies that are positive for "shish stain; and (3) selecting the implanted bacterial colony by using an inoculating loop, performing line drawing purification again on an MRS solid culture medium plate, performing purification twice, selecting the bacterial colony on the purified bacterial plate, inoculating the bacterial colony in an MRS liquid culture medium, performing amplification culture, and selecting the bacterial colony after culturing for 24 hours.
The bacteria are sent to the China center for type culture Collection of Wuhan university for strain preservation, and the strains are determined as follows:
candida parapsilosis (Candida humils) GS2 was deposited in the China, Wuhan university, China center for type culture Collection in 2018, month 07, month 02, with a collection number of CCTCC M2018441;
lactobacillus plantarum GS3 was deposited in China, Wuhan university, China center for type culture Collection in 2018, 08-03 month, with a collection number of CCTCC M2018522;
lactobacillus casei (Lactobacillus casei) GS4 was deposited in China, Wuhan university, China center for type culture Collection in 2018, 08-03 month, with a collection number of CCTCC M2018523;
lactobacillus paracasei (Lactobacillus paracasei) GS5 was deposited in China, Wuhan university, China center for type culture Collection in 2018, 08-03.month with the collection number of CCTCC M2018524.
Example 2, preparation of complex microbial inoculum:
1. materials and methods
2.1 test materials
1.1.1 conventional materials: fresh sugarcane tails, elephant grass, trehalose, skimmed milk powder, glycerol, sodium glutamate, urea and the like.
1.1.2 microbial strains: candida parapsilosis (Candida humils), Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus casei (Lactobacillus casei) and Lactobacillus paracasei (Lactobacillus paracasei).
1.1.3 Medium: MRS liquid culture medium is used for lactobacillus activation and bacterial liquid culture; MRS solid culture medium is used for lactobacillus plating counting; YPD liquid culture medium is used for yeast activation and bacterial liquid culture; YPD agar was used for yeast plating.
1.1.4 protective agent:
10% of skimmed milk powder, 8% of trehalose, 7.5% of sodium glutamate and 4% of glycerol.
1.2 test methods
1.2.1 formulation of Compound silage inoculant specially used for sugarcane tails
1.2.1.1 the addition of each strain is selected to determine different complex microbial inoculum formulas, and the dosage in the formula table is measured by viable count.
TABLE 1 sugarcane tail silage composite microbial inoculum formula
Unit: is based on
1.2.1.2 preparation of Complex microbial inoculum
The preparation process of the composite microbial inoculum comprises the following steps:
1.2.1.2.1 bacteria activation and culture
And (3) activation: inoculating 10% of the extracted strain into a liquid culture medium, and performing shake culture at 150r/min at 28 deg.C for 24 h; the lactobacillus culture temperature is 37 ℃ for 24h, and the lactobacillus is statically cultured.
And (3) amplification culture: inoculating 5% of the yeast into liquid culture medium, and performing shake culture at the culture temperature of 28 deg.C for 24 hr at 150 r/min; the lactobacillus culture temperature is 37 ℃ for 24h, and the lactobacillus is statically cultured.
The inoculated bacteria solution was counted with a smear.
1.2.1.2.2 Freeze drying
Mixing the yeast and the lactic acid bacteria after the concentrated culture according to the concentration of the bacteria and the proportion of an experimental scheme, and then carrying out a freeze drying test.
Preparation of bacterial suspension:
centrifuging the concentrated and cultured bacteria fermentation liquor at 3000r/min, sterilizing the centrifugal tube in advance, wiping the inner wall of the centrifugal machine with 95% ethanol, discarding supernatant, and mixing with appropriate amount of sterile water and bacteria mud to obtain bacteria suspension with uniform concentration.
Treating the protective agent:
pretreatment of a protective agent: and (4) ultraviolet sterilization is carried out for 2h, so that the sterility in the protective agent is ensured, and the influence on the test is avoided.
Pre-treatment of freeze drying:
adding a protective agent in the process of preparing the bacterial suspension, wherein the bacterial sludge: and (3) transferring the protective agents into a sterile culture dish together with a ratio of 1:3, wrapping the protective agents with a preservative film, placing the protective agents in an ultralow temperature refrigerator in parallel, pre-freezing the protective agents for 5 hours to form small ice crystals, immediately transferring the protective agents into a freeze drying device for drying until no clear water exists, taking out the protective agents, crushing the protective agents, sieving the protective agents with a 200-mesh sieve, sampling and measuring the number of viable bacteria, immediately performing vacuum sealing packaging, and then storing the protective agents with a 4-degree refrigerator.
1.2.2 sugarcane Tail silage fermentation test
The single comparison experiments using the bacterial agents of Lactobacillus brevis (Lactobacillus plantarum), Lactobacillus casei (Lactobacillus casei), Lactobacillus paracasei (Lactobacillus paracasei) and Candida albicans (Candida humils) as the leavening agents respectively and independently are formulas 1, 2, 3 and 4.
The blank control group is selected as a control group by adding normal saline and not adding any leavening agent.
In addition, on the basis of a self-made compound bacterial agent product, the addition amount of each bacterial liquid is 10ml, fresh sugarcane tail leaves cut into 2-3cm are respectively added, 3 times of each group are repeated, and all prepared ensilage bags are respectively marked as formulas 5 and 6.
The addition amount of the bacterial liquid or the normal saline corresponding to each silage bag is 10 ml.
2. Ensiling quality inspection
The bagged silage is stored in a shade place, and is subjected to unpacking inspection and evaluation at day 60, the smell, color and texture of the silage sample are observed and recorded (executed according to an attached table), and the sample is collected.
2.1 items and methods of measurement
(1) Sensory evaluation
According to the attached table item by item.
Sensory evaluation was performed with reference to "silage quality evaluation standards (trial) released in 1996 by the ministry of agriculture in China, and is specifically shown in table 3.
TABLE 2 sensory evaluation criteria
TABLE 3 sensory evaluation criteria
| Group of | Smell(s) | Color | Texture of |
| Control group | 17 | 17 | 8 |
| Formulation 1 | 20.5 | 17.5 | 9 |
| Formulation 2 | 20 | 18 | 8.5 |
| Formulation 3 | 20.5 | 18 | 8.5 |
| Formulation 4 | 19 | 17.5 | 8 |
| Formulation 5 | 23.5 | 19.5 | 9.5 |
| Formulation 6 | 20 | 18 | 9 |
Therefore, after the sugarcane tail silage is finished, the bags are opened, sensory evaluation scoring is respectively carried out on the three aspects of color, smell and texture, and the sensory evaluation of each treatment group is superior in quality. The silage quality evaluation grades of the sugarcane tail silage are good or superior grades by various silage laboratory evaluation methods. The total score of other laboratory silage quality evaluation methods adopted by each test group is higher than that of a control group, and the results show that lactobacillus plantarum, lactobacillus casei, lactobacillus paracasei and candida parapsilosis have a certain improvement effect on fermented sugarcane tail leaves. The silage indexes evaluated by the silage quality evaluation methods are different, so that the total scores of the test groups are different.
(2) Conventional nutrient content determination
DM, CP, NDF, ADF, Total energy, and ash content in the silage sugarcane tails were determined.
(3) Analysis of pH and quality items
(1) Sample pretreatment
Taking 35g of sample per bag, placing into 250mL wide-mouth bottle, adding 150mL of deionized water, placing in 4 deg.C refrigerator for extraction for 24 hr, filtering with 2 layers of gauze, and using the filtrate for pH, lactic acid, and ammoniacal Nitrogen (NH)3-N) and Volatile Fatty Acids (VFA). And (3) drying and crushing the residual sugarcane tail sample at 65 ℃ except the sugarcane tail used for the screening and culturing of the germ-removing seeds (sieving with a 40-mesh sieve) to prepare an air-dried sample for measuring the conventional nutrient components.
(2) Determination of pH
And (3) measuring by adopting a pH meter, wherein the type of the pH meter is as follows: HANNA HI 8424.
(3) The lactate determination principle takes NAD + as a hydrogen acceptor, and LDH catalyzes lactate dehydrogenation to generate pyruvic acid, so that NAD + is converted into NADH. Wherein the hydrogen transfer of PMS reduces NBT into purple color substance, and the absorbance of the color substance is in linear relation with the content of lactic acid when the absorbance is 530 nm.
(4)NH3Determination of-N
NH3N is measured by a colorimetric method, 5mL of the obtained filtrate is taken, 5mL of 0.2mol/L HCL of 5ML is added, and 3500rpm is centrifuged for 10min to take the supernatant. Using ammonium chloride as standard, performing color comparison at 700nm with ultraviolet-visible spectrophotometer (PE Lambda 35 type, produced in USA), and obtaining NH according to optical density value and standard curve3-the content of N.
(5) Determination of VFA
Taking 0.5mL of obtained filtrate, adding 0.5mL of 8.2% metaphosphoric acid, centrifuging at 13000rpm/min for 10min, taking supernatant, adding crotonic acid, and measuring by an Agilent 7890A type gas chromatograph, wherein a capillary column is adopted: HP-INNOWAX (19091N-133), column Specification of capillary: 30m multiplied by 0.25mm multiplied by 0.25um, and the contents of the B, C and butyric acids of the samples are measured.
(6) Determination of ethanol
2mL of the filtrate is taken and centrifuged for 15min in a 2mL centrifuge tube 12000rpm/min, and the supernatant is taken and filtered through a 0.45um microporous filter membrane. Taking 0.9mL of the treated filtrate, adding 0.1mL of n-propanol into the GC bottle, uniformly mixing, measuring by an Agilent 7890A type gas chromatograph, and carrying out capillary column: HP-INNOWAX (19091N-133), column Specification of capillary: 30 m.times.0.25 mm.times.0.25 um, and the ethanol content of the sample was measured.
2.2 data analysis
Data were initially processed using EXCEL software, single factor analysis of variance using SPSS 16.0 software, and Duncan multiple comparisons.
2.3 results and analysis
TABLE 4 influence on fermentation quality of sugarcane Tail silage
From the table, the feed prepared by fully mixing silage with the compound agent of the formula 5 can effectively improve the indexes of lactic acid and acetic acid and obviously reduce the content of ammoniacal nitrogen in silage (P is less than 0.05).
TABLE 5 influence of composite micro-ecological agents on the quality of sugarcane tail mixed silage nutrition (absolutely dry basis)
As can be seen from the above table, the NDF and ADF of the mixed bacteria of formula 5 are reduced compared with those of the blank group and formulas 1-4.
Although the present invention has been described with respect to the preferred embodiments, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (6)
1. The composite microbial inoculum for ensiling the sugarcane tail leaves is characterized in that: its active component includes candida parapsilosis (A)Candida humils) GS2, Lactobacillus plantarum (C.) (L.) (Lactobacillus plantarum) GS3, Lactobacillus casei (L.casei) ((L.casei))Lactobacillus casei) GS4, Lactobacillus paracasei (L.) (Lactobacillus paracasei）GS5；
The Candida parapsilosis (C.), (Candida humils) GS2 was deposited in China, Wuhan university, China center for type culture Collection in 2018, 7/02/month, with a deposition number of CCTCC NO: m2018441;
lactobacillus plantarum GS3 was deposited in China, Wuhan university, China center for type culture Collection in 2018 at 08 month 03 with a deposition number of CCTCC NO: m2018522;
lactobacillus casei (Lactobacillus casei) GS4 was deposited in china, wuhan, university of wuhan, china type culture collection, in 2018 at 03 month 08 with a deposition number of CCTCC NO: m2018523;
lactobacillus paracasei (Lactobacillus paracasei) GS5 was deposited in china, wuhan, university of wuhan, china type culture collection, in 2018 at month 08 and 03 with a deposition number of CCTCC NO: m2018524;
the Candida parapsilosis (C.), (Candida humils) GS2, Lactobacillus plantarum (C.) (L.) (Lactobacillus plantarum) GS3, Lactobacillus casei (L.casei) ((L.casei))Lactobacillus casei) GS4, Lactobacillus paracasei (L.) (Lactobacillus paracasei) The percentage of GS5 is 20 percent to 60 percent to 10 percent in terms of viable count.
2. The preparation method of the composite microbial inoculum for silage of sugarcane tail leaves as claimed in claim 1, which is characterized in that: the method specifically comprises the following steps:
(1) and activating: extracting 4 kinds of bacteria, inoculating to liquid culture medium, and inoculating to Candida parapsilosis (Candida parapsilosis) in an amount of 10%Candida humils) Culturing GS2 at 28 deg.C for 24h, and shake culturing at 150 r/min; lactobacillus plantarum (A)Lactobacillus plantarumGS 3), Lactobacillus casei (Lactobacillus casei)Lactobacillus casei) GS4, Lactobacillus paracasei (L.) (Lactobacillus paracasei) GS5 is cultured statically at 37 ℃ for 24 h;
(2) and (3) expanding culture: then respectively inoculating the activated 4 strains into a liquid culture medium and candida parapsilosis (Candida parapsilosis) according to the addition amount of 5 percentCandida humils) Culturing GS2 at 28 deg.C for 24h, and shake culturing at 150 r/min; lactobacillus plantarum (A)Lactobacillus plantarum) GS3, Lactobacillus casei (L.casei) ((L.casei))Lactobacillus casei) GS4, Lactobacillus paracasei (L.) (Lactobacillus paracasei) GS5 culture temperature is 37 ℃ for 24h, static culture, and plate coating is used for counting inoculated bacteria liquid;
(3) and (3) freeze drying: wiping the inner wall of a centrifuge with 95% ethanol to perform sterilization treatment, mixing concentrated and cultured bacteria fermentation liquor according to the concentration proportion of 4 strains by using a sterilization centrifugal tube, centrifuging for 20min at 3000r/min, discarding supernatant to obtain bacteria mud, fully mixing the bacteria mud with a proper amount of sterile water to prepare bacteria suspension with uniform concentration, and adding a protective agent, namely the bacteria mud: and (3) the protective agent =1:3, the mixture is transferred into a sterile culture dish, the thickness of the protective agent is half of the height of the dish, the protective agent is wrapped by a preservative film, the dish is placed in an ultralow temperature refrigerator for pre-freezing for 5 hours to form small ice crystals, the ice crystals are immediately transferred into a freeze drying device for drying until no clear water exists, the number of viable bacteria at the moment is measured by sampling, and the mixture is immediately subjected to vacuum sealing packaging and then is stored in the refrigerator at 4 ℃.
3. The method of claim 2, wherein: the protective agent comprises, by percentage, 80% of fat-free milk powder, 8% of trehalose, 8% of sodium glutamate and 4% of glycerol.
4. The method of claim 2, wherein: in steps (1) to (2), MRS liquid medium is used for Lactobacillus plantarum ((R))Lactobacillus plantarum) GS3, Lactobacillus casei (L.casei) ((L.casei))Lactobacillus casei) GS4, Lactobacillus paracasei (L.) (Lactobacillus paracasei) Activating GS5 and culturing bacterial liquid; MRS solid culture medium for lactobacillus plantarum (MRS)Lactobacillus plantarum) GS3, Lactobacillus casei (L.casei) ((L.casei))Lactobacillus casei) GS4, Lactobacillus paracasei (L.) (Lactobacillus paracasei) GS5 plate coating count; YPD liquid culture medium for Candida parapsilosis (YPD)Candida humils) Activating GS2 and culturing bacterial liquid; YPD agar for Candida parapsilosis (YPD agar)Candida humils) GS2 plate count.
5. The use of the complex microbial inoculant for the silage of sugarcane tail leaves according to any one of claims 1 to 4 in the preparation of the silage of sugarcane tail leaves.
6. The use method of the composite bacteria for the silage of sugarcane tail leaves as claimed in claim 1, characterized in that: comprises the following steps: cutting fresh sugarcane tail leaves into 2-3cm, spraying the composite microbial inoculum for ensiling the sugarcane tail leaves on the fresh sugarcane tail leaves, uniformly mixing, loading into an ensiling bag, vacuumizing, and fermenting for 60 days.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911305751.3A CN111304112B (en) | 2019-12-12 | 2019-12-12 | Composite microbial inoculum for ensiling sugarcane tail leaves |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911305751.3A CN111304112B (en) | 2019-12-12 | 2019-12-12 | Composite microbial inoculum for ensiling sugarcane tail leaves |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111304112A CN111304112A (en) | 2020-06-19 |
| CN111304112B true CN111304112B (en) | 2021-10-01 |
Family
ID=71156385
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911305751.3A Active CN111304112B (en) | 2019-12-12 | 2019-12-12 | Composite microbial inoculum for ensiling sugarcane tail leaves |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111304112B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113995052B (en) * | 2021-07-12 | 2023-11-17 | 广西壮族自治区水牛研究所 | Ivory sugarcane tail leaf silage and preparation method thereof |
| CN114276949B (en) * | 2021-11-10 | 2023-05-05 | 丰唐生态农业科技研发(山东)有限公司 | Lactobacillus paracasei and application thereof in preparing silage with folium Broussonetiae |
| CN117778278B (en) * | 2024-02-27 | 2024-05-03 | 山东中科嘉亿生物工程有限公司 | Compound lactobacillus silage additive and preparation method and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108949629A (en) * | 2018-07-21 | 2018-12-07 | 四川农业大学 | A kind of composite ferment and the preparation method and application thereof |
-
2019
- 2019-12-12 CN CN201911305751.3A patent/CN111304112B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108949629A (en) * | 2018-07-21 | 2018-12-07 | 四川农业大学 | A kind of composite ferment and the preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111304112A (en) | 2020-06-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111304112B (en) | Composite microbial inoculum for ensiling sugarcane tail leaves | |
| CN107047978A (en) | A kind of Lactobacillus plantarum and its application in ensilage is prepared | |
| CN102643767B (en) | Lactobacillus plantarum and application thereof in fermenting and ensiling sweet potato stem and leaf | |
| CN111944731A (en) | Method for improving flavor of fermented fruits and vegetables based on multi-bacterium co-fermentation technology | |
| CN109837217B (en) | High-temperature-resistant Pleurotus ostreatus and screening method and application thereof | |
| CN102093965B (en) | Lactic acid bacteria and application thereof | |
| CN109504614A (en) | A kind of preparation method of native country saccharomyces cerevisiae dry powder | |
| CN108060102A (en) | A kind of cibarium Wei Si Salmonellas and its application, screening calibration method | |
| CN110982739B (en) | Lactobacillus plantarum for high-moisture oat silage and application thereof | |
| CN113151032A (en) | Bacillus subtilis with efficient gossypol degradation capability and application thereof | |
| CN110591932B (en) | Yeast MA for controlling postharvest diseases of fruits and vegetables and use method thereof | |
| CN108060103A (en) | A kind of Pediococcus pentosaceus and its application, screening calibration method | |
| CN110331104B (en) | Lactobacillus plantarum CV10D1 and application thereof | |
| CN112251388A (en) | Lactobacillus plantarum and application of lactobacillus leavening agent thereof | |
| CN111748490A (en) | Lactobacillus sake and application thereof | |
| CN113133498A (en) | Composite microbial additive for improving quality of silage and application thereof | |
| CN114908018B (en) | Lactobacillus brevis and application thereof in silage | |
| CN115011515B (en) | Strain for increasing plant alcohol content in alfalfa silage and application thereof | |
| CN106615609B (en) | Lactobacillus buchneri capable of improving silage quality and application thereof | |
| CN113736671B (en) | Endophytic fungi AM8.2 for promoting potassium element absorption of camellia oleifera | |
| CN102640886B (en) | Applications of Lactobacillus plantarum in Brassica oleracea outer wrapper leaf silage | |
| CN111096392A (en) | Full-mixed silage of sugarcane tail leaves and application thereof | |
| CN112522119A (en) | Preparation method and application of combined microbial inoculum based on Klebsiella citrullina and Candida intermedia | |
| CN117660250B (en) | Low-temperature lactobacillus and application thereof | |
| CN113768045B (en) | Mixed silage method for sweet potato peels and tea residues |
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 | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20200619 Assignee: Guangxi Yulin Yiliang Technology Co.,Ltd. Assignor: GUANGXI ZHUANG AUTONOMOUS REGION BUFFALO INSTITUTE Contract record no.: X2023980045755 Denomination of invention: A composite bacterial agent for sugarcane tail leaf silage Granted publication date: 20211001 License type: Common License Record date: 20231107 |
|
| EE01 | Entry into force of recordation of patent licensing contract |