CN110129233A - A kind of microbial bacterial agent and preparation method thereof being used to prepare cage bird fermenting bed padding - Google Patents
A kind of microbial bacterial agent and preparation method thereof being used to prepare cage bird fermenting bed padding Download PDFInfo
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- CN110129233A CN110129233A CN201910431959.3A CN201910431959A CN110129233A CN 110129233 A CN110129233 A CN 110129233A CN 201910431959 A CN201910431959 A CN 201910431959A CN 110129233 A CN110129233 A CN 110129233A
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- padding
- fermentation
- microbial
- organism fungicide
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- 230000000813 microbial effect Effects 0.000 title claims abstract description 38
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 230000001580 bacterial effect Effects 0.000 title abstract description 17
- 238000000855 fermentation Methods 0.000 claims abstract description 53
- 230000004151 fermentation Effects 0.000 claims abstract description 53
- 241000894006 Bacteria Species 0.000 claims abstract description 42
- 244000005700 microbiome Species 0.000 claims abstract description 38
- 230000000855 fungicidal effect Effects 0.000 claims abstract description 27
- 239000000417 fungicide Substances 0.000 claims abstract description 27
- 239000007787 solid Substances 0.000 claims abstract description 22
- 241000194107 Bacillus megaterium Species 0.000 claims abstract description 21
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 20
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 19
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 19
- 239000007788 liquid Substances 0.000 claims abstract description 16
- 241000193744 Bacillus amyloliquefaciens Species 0.000 claims abstract description 13
- 241000223259 Trichoderma Species 0.000 claims abstract description 12
- 238000002156 mixing Methods 0.000 claims abstract description 12
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 11
- 238000001035 drying Methods 0.000 claims abstract description 11
- 239000000203 mixture Substances 0.000 claims abstract description 9
- 239000002131 composite material Substances 0.000 claims description 22
- 235000015099 wheat brans Nutrition 0.000 claims description 15
- 240000007594 Oryza sativa Species 0.000 claims description 9
- 235000007164 Oryza sativa Nutrition 0.000 claims description 9
- 239000010903 husk Substances 0.000 claims description 9
- 239000002068 microbial inoculum Substances 0.000 claims description 9
- 235000009566 rice Nutrition 0.000 claims description 9
- 235000001674 Agaricus brunnescens Nutrition 0.000 claims description 8
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 6
- 239000002689 soil Substances 0.000 claims description 6
- 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 claims description 5
- 244000068988 Glycine max Species 0.000 claims description 5
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- 239000010871 livestock manure Substances 0.000 abstract description 19
- 230000015556 catabolic process Effects 0.000 abstract description 11
- 238000006731 degradation reaction Methods 0.000 abstract description 11
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 abstract description 8
- 230000012010 growth Effects 0.000 abstract description 8
- 239000005416 organic matter Substances 0.000 abstract description 7
- 229910021529 ammonia Inorganic materials 0.000 abstract description 4
- 241000894007 species Species 0.000 abstract description 3
- 230000000295 complement effect Effects 0.000 abstract description 2
- 238000004332 deodorization Methods 0.000 abstract description 2
- 201000010099 disease Diseases 0.000 abstract description 2
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- 239000012530 fluid Substances 0.000 abstract 1
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 26
- 239000001963 growth medium Substances 0.000 description 13
- 239000002609 medium Substances 0.000 description 13
- 229910052757 nitrogen Inorganic materials 0.000 description 13
- 241000193830 Bacillus <bacterium> Species 0.000 description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 10
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 10
- 238000000034 method Methods 0.000 description 10
- 238000004321 preservation Methods 0.000 description 9
- 241000726221 Gemma Species 0.000 description 7
- 238000004519 manufacturing process Methods 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- 239000002253 acid Substances 0.000 description 6
- 239000003864 humus Substances 0.000 description 5
- 239000004310 lactic acid Substances 0.000 description 5
- 235000014655 lactic acid Nutrition 0.000 description 5
- 239000003895 organic fertilizer Substances 0.000 description 5
- 239000011780 sodium chloride Substances 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- QJZYHAIUNVAGQP-UHFFFAOYSA-N 3-nitrobicyclo[2.2.1]hept-5-ene-2,3-dicarboxylic acid Chemical compound C1C2C=CC1C(C(=O)O)C2(C(O)=O)[N+]([O-])=O QJZYHAIUNVAGQP-UHFFFAOYSA-N 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 4
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 239000004021 humic acid Substances 0.000 description 4
- 239000006916 nutrient agar Substances 0.000 description 4
- KMUONIBRACKNSN-UHFFFAOYSA-N potassium dichromate Chemical compound [K+].[K+].[O-][Cr](=O)(=O)O[Cr]([O-])(=O)=O KMUONIBRACKNSN-UHFFFAOYSA-N 0.000 description 4
- 238000011218 seed culture Methods 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- 102000016938 Catalase Human genes 0.000 description 3
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- 241000191967 Staphylococcus aureus Species 0.000 description 3
- 229920002472 Starch Polymers 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- 239000004382 Amylase Substances 0.000 description 2
- 102000013142 Amylases Human genes 0.000 description 2
- 108010065511 Amylases Proteins 0.000 description 2
- 241000589152 Azotobacter chroococcum Species 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- 206010059410 Faecaluria Diseases 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 241000108664 Nitrobacteria Species 0.000 description 2
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- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 2
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 2
- 238000010564 aerobic fermentation Methods 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 2
- 235000011130 ammonium sulphate Nutrition 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 235000019418 amylase Nutrition 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000004737 colorimetric analysis Methods 0.000 description 2
- 239000002361 compost Substances 0.000 description 2
- 239000012531 culture fluid Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000006153 eosin methylene blue Substances 0.000 description 2
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- 238000010438 heat treatment Methods 0.000 description 2
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- 229910052698 phosphorus Inorganic materials 0.000 description 2
- 239000011574 phosphorus Substances 0.000 description 2
- 239000011591 potassium Substances 0.000 description 2
- 229910052700 potassium Inorganic materials 0.000 description 2
- 239000001103 potassium chloride Substances 0.000 description 2
- 235000011164 potassium chloride Nutrition 0.000 description 2
- 239000001965 potato dextrose agar Substances 0.000 description 2
- 244000144977 poultry Species 0.000 description 2
- 230000008569 process Effects 0.000 description 2
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- 238000011160 research Methods 0.000 description 2
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- 239000002023 wood Substances 0.000 description 2
- FFRBMBIXVSCUFS-UHFFFAOYSA-N 2,4-dinitro-1-naphthol Chemical compound C1=CC=C2C(O)=C([N+]([O-])=O)C=C([N+]([O-])=O)C2=C1 FFRBMBIXVSCUFS-UHFFFAOYSA-N 0.000 description 1
- 241000609240 Ambelania acida Species 0.000 description 1
- 108010059892 Cellulase Proteins 0.000 description 1
- 241001478240 Coccus Species 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
- 241000192125 Firmicutes Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000736262 Microbiota Species 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 1
- 241001072230 Oceanobacillus Species 0.000 description 1
- 108091006629 SLC13A2 Proteins 0.000 description 1
- 235000009754 Vitis X bourquina Nutrition 0.000 description 1
- 235000012333 Vitis X labruscana Nutrition 0.000 description 1
- 240000006365 Vitis vinifera Species 0.000 description 1
- 235000014787 Vitis vinifera Nutrition 0.000 description 1
- 241000817483 Vulgatibacter Species 0.000 description 1
- FRYDSOYOHWGSMD-UHFFFAOYSA-N [C].O Chemical compound [C].O FRYDSOYOHWGSMD-UHFFFAOYSA-N 0.000 description 1
- JPONAEJXLMLEKR-UHFFFAOYSA-L [OH-].[Na+].[O-]P(O)(=O)OP(=O)(O)O.[Na+] Chemical compound [OH-].[Na+].[O-]P(O)(=O)OP(=O)(O)O.[Na+] JPONAEJXLMLEKR-UHFFFAOYSA-L 0.000 description 1
- 238000003915 air pollution Methods 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 239000010905 bagasse Substances 0.000 description 1
- 239000007640 basal medium Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 229940106157 cellulase Drugs 0.000 description 1
- 238000009642 citrate test Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- DMBHHRLKUKUOEG-UHFFFAOYSA-N diphenylamine Chemical class C=1C=CC=CC=1NC1=CC=CC=C1 DMBHHRLKUKUOEG-UHFFFAOYSA-N 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- CEQFOVLGLXCDCX-WUKNDPDISA-N methyl red Chemical compound C1=CC(N(C)C)=CC=C1\N=N\C1=CC=CC=C1C(O)=O CEQFOVLGLXCDCX-WUKNDPDISA-N 0.000 description 1
- HPNSFSBZBAHARI-UHFFFAOYSA-N micophenolic acid Natural products OC1=C(CC=C(C)CCC(O)=O)C(OC)=C(C)C2=C1C(=O)OC2 HPNSFSBZBAHARI-UHFFFAOYSA-N 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 229940014456 mycophenolate Drugs 0.000 description 1
- HPNSFSBZBAHARI-RUDMXATFSA-N mycophenolic acid Chemical compound OC1=C(C\C=C(/C)CCC(O)=O)C(OC)=C(C)C2=C1C(=O)OC2 HPNSFSBZBAHARI-RUDMXATFSA-N 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- DHXDPKBQYJXIDQ-UHFFFAOYSA-M potassium sodium 3-carboxy-3,5-dihydroxy-5-oxopentanoate nitrate Chemical compound [N+](=O)([O-])[O-].[K+].C(CC(O)(C(=O)O)CC(=O)O)(=O)[O-].[Na+] DHXDPKBQYJXIDQ-UHFFFAOYSA-M 0.000 description 1
- 238000009374 poultry farming Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000010902 straw Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K31/00—Housing birds
- A01K31/04—Dropping-boards; Devices for removing excrement
-
- 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
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Environmental Sciences (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Mycology (AREA)
- Botany (AREA)
- Birds (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a kind of microbial bacterial agents and preparation method thereof for being used to prepare cage bird fermenting bed padding.The complex micro organism fungicide, be by bacillus amyloliquefaciens fermentation liquid, bacillus megaterium fermentation liquid, long shoot trichoderma fermentation liquid and lactobacillus plantarum zymotic fluid group at composition be inoculated in solid medium, mixing, which is collectively disposed under 30-35 DEG C of condition of culture, cultivates 72-96h, and drying is made.Complex micro organism fungicide of the present invention is preparing the application in cage bird fermenting bed padding.The present invention is prepared into complex micro organism fungicide with four kinds of excellent species selection optimal proportions, function stem acts synergistically during the feeding management of cage bird fermentation bed surface, organic matter transformation bacterium, solid ammonia deodorization bacterium and germ inhibit bacterium to have complementary functions, the fermentation at elevated temperatures of quick start fermentation bed, it realizes the fast eliminating of moisture in excrement, improves fermentation bed microbial diversity, inhibit the growth of cause of disease spoilage organisms, promote chicken manure ira situ degradation, improves padding humic rate.
Description
Technical field
The invention discloses a kind of microbial bacterial agents and preparation method thereof for being used to prepare cage bird fermenting bed padding, belong to
Microbial engineering field.
Background technique
Currently, scale chicken house uses the yard feeding pattern of cage, cage mode is in production level, the labor efficiency for improving chicken house
While, it concentrates and discharges there is also chicken manure, the problems such as chicken house itself environmental degradation, fecal pollution, which has become, to be restricted cage bird and support
An important factor for growing.The fecaluria processing technique of cage bird mostly uses greatly compost fermentation, biogas fermentation, aerobic fermentation drying technology etc.
Mode is handled, and compost fermentation handles excrement land occupation, larger to the air pollution of environment, and biogas fermentation has season office
Sex-limited, aerobic fermentation drying technology higher cost has certain limitation in actually utilizing.
Fermentation bed cultivation technology is a kind of novel ecological aquaculture model, and key is to be laid with fermentation pad on livestock and poultry colony house ground
Material, realizes the ira situ degradation of Animal fecal pollution, greatly improves breeding environment, is an environmentally friendly agricultural technology.Microbial fermentation
Bed mode realize cultivation, excrement in situ decomposed fermentation stage synchronize, be the important channel of rapid production of organic fertilizer.Humus
It is the important component of organic fertilizer, humic acid fertilizer addition is remarkably improved soil fertility, promotes plant growth.Fermentation bed master at present
To be applied to the cultivation of live pig, the research for poultry farming is less, and the fermentation bed research about cage bird is also rarely reported.
Tradition poultry padding is relatively thin, and generally in 10cm or so, variation of the chicken manure in padding does not have based on decay process
There is Fermentation Function, little to chicken manure decomposition, padding mainly acts to adsorb chicken manure moisture and foul smell, unobvious heat production,
Padding is wetter, and foul smell is serious in padding and henhouse.The fermenting bed padding thickness of cage bird is different from traditional flat feeding padding mould
Formula, the thickness and proportion of padding significantly affect the porosity and retentiveness of fermentation padding.Pad material fermentation decompose fecaluria process be
Microbial action as a result, the height of fermenting microbe vigor determine excrement decompose and humus generate efficiency, be microorganism
The core link of deep-litter systems.Selection colonization ability is strong, degradation rate is high and function is answered with what the bacterial strain of complementation formed
Microbial bacterial agent is closed, is the advantageous guarantee that fermentation bed ira situ degradation excrement forms humic acid organic fertilizer.
Summary of the invention
The purpose of the present invention is intended to provide one kind and effectively facilitates chicken manure ira situ degradation, significantly reduces padding ammoniacal nitrogen and aqueous
Amount improves the microbial bacterial agent and preparation method thereof of the cage bird fermentation bed of padding humic rate.
The purpose of the present invention can be achieved through the following technical solutions:
A kind of microbial composite being used to prepare cage bird fermenting bed padding, the microbial fermentation matched by following volumes
Liquid composition: bacillus amyloliquefaciens fermentation liquid 20-30%, bacillus megaterium fermentation liquid 25-35%, long shoot trichoderma fermentation liquid
20-30%, lactobacillus plantarum fermentation liquid 15-25%, total viable count contained by microbial inoculum is between 4 × 109-8 × 109cfu/g.
The microbial composite is preferably made of the microbial fermentation solution that following volumes matches: solution starch gemma bar
Bacterium 25%, bacillus megaterium 30%, long shoot trichoderma 25%, lactobacillus plantarum 20%, total viable count contained by microbial inoculum 4 ×
109-8×109Between cfu/g.
1, the selection of excellent species
Moisture content 50% in fresh chicken manure, organic matter 25.5%, nitrogen 1.63%, phosphorus 1.54%, potassium 0.85%, carbon hydrate
Object 11%, fiber 7%, and moisture content 60-65% in pig manure, organic matter 15%, nitrogen 0.5%, phosphorus 0.5~0.6%, potassium 0.35~
0.45%.The organic matter and nitrogen content of chicken manure are all relatively high, therefore should select that protease, amylase can be produced, and convert ammoniacal nitrogen
Strain collocation cellulase producing strain, produce the antibacterial bacterial strain of acid, reduce the ammoniacal nitrogen in excrement conversion, conversion excrement is organic
Matter improves padding humus.
Bacillus amyloliquefaciens (CICC 20164), produce protease, amylase, can utilize rapidly albumen, fat, carbohydrate,
The organic carbons such as starch promote the conversion of degradable object in matrix in a short time, accelerate the heating of matrix padding.
Long shoot trichoderma (CICC 41185), it breeds rapidly, in a short time fast-growth, metabolism, has relatively strong decompose
It is bent to produce cellulose mycophenolate using bagasse, stalk, mushroom bran as substrate for the ability of cellulose.
Bacillus megatherium, the bacillus azotobacter with nitrification screened from chicken manure, the bacterial strain have been preserved in
China typical culture collection center, preservation address: Bayi Road No. 299 Wuhan Universitys in Wuhan City, Hubei Province Wuchang District in the school, divide
Class name: bacillus megaterium Bacillus megaterium, deposit number: CCTCC No:M2019050, preservation date are as follows:
On January 30th, 2019.Bacterium colony surface is smooth opaque, and in dirty white, cell is elongated rod shape, and gram-positive bacteria is trained at 37 DEG C
2~3 days formation gemma are supported, 0.6~1.5 μm of gemma size, gemma is oval.
Lactobacillus plantarum is that acid production speed is fast, growth ability is strong, inhibits pathogen for having of screening from stalk
Lactic acid bacteria；The bacterial strain has been preserved in China typical culture collection center, preservation address: Wuhan City, Hubei Province Wuchang District Bayi Road
No. 299 Wuhan Universitys in the school, classification naming: lactobacillus plantarum Lactobacillus plantarum, deposit number: CCTCC No:
M2016281, preservation date are as follows: on May 25th, 2016.The bacterium colony of lactobacillus plantarum is creamy white, neat in edge；Cell is micro-
Form is rod-short, no gemma, Gram-positive；Catalase is negative, oxidase negative；With stronger growth and production acid
Performance can grow on the culture medium that initial ph value is 3.0-8.0, faint growth be presented under the conditions of 10 DEG C, at 15 DEG C -45 DEG C
Under the conditions of upgrowth situation it is good, be not more than 6.5%NaCl under conditions of upgrowth situation it is good.
The preparation of microbial composite of the present invention:
1, level-one spreads cultivation: the strain of each purified culture (bacillus amyloliquefaciens, bacillus megaterium, long shoot trichoderma,
Lactobacillus plantarum) it after taking-up, is inoculated into five seed culture mediums prepared and cultivates respectively respectively, often from the inclined-plane of preservation
Culture medium 200ml is filled in the bottle of a 1L, lactobacillus plantarum cultivates 48h in 37 DEG C of constant incubator, bacillus amyloliquefaciens,
Bacillus megaterium 35 DEG C of 150r/min shaken cultivation 48h on shaking table, long shoot trichoderma 30 DEG C of 120r/min oscillations on shaking table
Cultivate 96h.
2, second level spreads cultivation: period detects each bacteria culture fluid viable count respectively, to each bacterium viable count >=2 × 108After/ml, take
Four kinds of bacterium the microbial composite are made by inoculating proportion by seed liquor out.
A kind of complex micro organism fungicide being used to prepare cage bird fermenting bed padding, the complex micro organism fungicide be by
Microbial composite of the present invention is inoculated in solid medium, and mixing, which is collectively disposed under 30-35 DEG C of condition of culture, cultivates
72-96h, drying are made.
The complex micro organism fungicide, preferably by microbial composite of the present invention and solid medium according to weight
Than being that microbial composite is inoculated in solid medium by the ratio of 1:9, mixing is collectively disposed under 30-35 DEG C of condition of culture amount
Cultivate 72-96h；Then drying can be made into complex micro organism fungicide under the conditions of 50 DEG C；The solid culture based formulas are as follows:
Soybean residue 64%, wheat bran 22%, corn flour 10%, glucose 3.5%, NaCl 0.4%, KH2PO40.1%, pH6.4-7.2.
The preparation method of complex micro organism fungicide of the present invention, it is solid including the microbial composite to be inoculated in
In body culture medium, mixing, which is collectively disposed under 30-35 DEG C of condition of culture, cultivates 72-96h, dries the composite microbial bacteria for being made described
Agent；It preferably includes for microbial composite of the present invention to be inoculated in solid medium, mixing is collectively disposed at 30-35 DEG C of training
72-96h is cultivated under the conditions of supporting；Then drying can be made into complex micro organism fungicide under the conditions of 50 DEG C.
The solid culture based formulas is preferably: soybean residue 64%, wheat bran 22%, corn flour 10%, glucose
3.5%, NaCl 0.4%, KH2PO40.1%, pH6.4-7.2；Microbial composite and solid medium of the present invention
Weight ratio is preferably 1:9.
Complex micro organism fungicide of the present invention is preparing the application in cage bird fermenting bed padding.
A kind of cage bird fermenting bed padding includes complex micro organism fungicide of the present invention.
It is also preferable to include mushroom bran, sawdust, rice husk and wheat brans for the cage bird fermenting bed padding.
The cage bird fermenting bed padding, is further preferably prepared via a method which: cage bird fermentation bed 35~
40cm mushroom bran, surface spread the weight ratios such as sawdust and rice husk 8~15cm of mixture, are run and are stirred with machine for turning up the soil and harrowing, by mushroom bran and sawdust
It is mixed with the mixture of rice husk；After mixing by complex micro organism fungicide, wheat bran described in claim 3 or 4, it is spread on fermentation
Bed surface, then run and stirred with machine for turning up the soil and harrowing, wherein；The weight ratio of complex micro organism fungicide of the present invention, wheat bran are as follows: 1:15
~22；Every square of padding spreads complex micro organism fungicide 50g of the present invention.
The beneficial effects of the present invention are:
(1) product of the present invention forms complex micro organism fungicide with four kinds of excellent species selection optimal proportions, and function stem exists
It can act synergistically during the feeding management of cage bird fermentation bed surface, organic matter transformation bacterium (bacillus amyloliquefaciens, long shoot wood
It is mould), solid ammonia deodorization bacterium (bacillus megaterium) and germ inhibition bacterium (lactobacillus plantarum) have complementary functions, quick start fermentation bed
Fermentation at elevated temperatures, realize excrement in moisture fast eliminating, improve fermentation bed microbial diversity, inhibit staphylococcus aureus,
The growth of the cause of diseases spoilage organisms such as popular Bacillus, effectively facilitates chicken manure ira situ degradation.
(2) bacillus megaterium separated from chicken manure can be colonized, fast-growth in cage bird fermentation bed, significant to drop
Low padding ammoniacal nitrogen reduces ammonia, while reducing nitrogen loss during heating.
(3) mushroom bran, rice husk, sawdust mat combination, cooperate complex micro organism fungicide, the humics such as humic acid can be significantly improved
The content of matter improves padding humic rate, forms high-quality organic fertilizer.
Detailed description of the invention
The flora abundance difference of Fig. 1 difference padding is analyzed
Biomaterial preservation information
Bacillus megatherium BM18, classification naming are Bacillus megaterium BM18, are preserved in Chinese Typical Representative training
Object collection is supported, deposit number: CCTCC No:M2019050, preservation address Wuhan, China Wuhan University, the deposit date is 2019
On January 15, in.Lactobacillus plantarum ZRR, classification naming are Lactobacillus plantarumZRR, are preserved in Chinese Typical Representative training
Object collection is supported, deposit number: CCTCC No:M2016281, preservation address Wuhan, China Wuhan University, the deposit date is 2016
On May 25, in.
Specific embodiment
Following implementation is intended to further illustrate the present invention, is not intended to limit the present invention.
The 4 plants of strain names and strain source that the present invention uses are as follows:
Bacillus amyloliquefaciens (CICC 20164), long shoot trichoderma (CICC 41185) are purchased from Chinese industrial microbial bacteria
Kind collection.Bacillus megatherium, the bacillus azotobacter with nitrification screened from chicken manure, has been preserved in China
Type Tissue Collection, deposit number: CCTCC No:M2019050.Lactobacillus plantarum is the tool screened from stalk
There is the lactic acid bacteria that acid production speed is fast, growth ability is strong, inhibits pathogen, be preserved in China typical culture collection center, protects
Hiding number: CCTCC No:M2016281.
Embodiment 1: the separation of bacillus megaterium and lactobacillus plantarum
The separation of bacillus megaterium: take the access of 10g Chicken dung equipped with 200mL enriched medium (ammonium sulfate 2g, amber
Sour sodium 2.17g, potassium dihydrogen phosphate 0.25g, magnesium sulfate 0.125g, NaC1 0.125g, water 1000mL) in, every 3d, with 3%
Inoculum concentration is transferred in new enriched medium, continuous enrichment culture 30d, on dilution spread to ammonium sulfate plate, selects growing way
Fast macrocolony, repeated isolation purify twice.By the strain inoculated of acquisition in enriched medium shaken cultivation, while with lattice benefit
This reagent, diphenylamines reagent tracing detection nitrite, nitrate production primary dcreening operation obtain 28 plants of bacterial strain, then bacterial strain is answered
It is connected in chicken manure leachate that (preparation of chicken manure leachate: chicken manure and tap water are mixed in l:1 ratio, 121 in high-pressure sterilizing pot
DEG C sterilizing 20min, left undisturbed overnight, filtering, again 12l DEG C of sterilizing 20min).It is surveyed using potassium chloride extraction-indophenol blue colorimetry
The content of fixed inoculation front and back ammoniacal nitrogen obtains strong 6 plants of bacterial strain of degradation capability, wherein one plant of bacterium 48 hours ammonia nitrogen degradation rates most
Height reaches 69.7%.The bacterial strain bacterium colony is larger, and edge is irregular；Thallus is in the shape of a rod, there is gemma, Gram-positive；Starch
Hydrolysis experiment, gelatin hydrolysis, oxidase test, catalase test, methyl red test, citrate test are positive, through 16rRNA
Gene order is accredited as bacillus megaterium, will be preserved in China typical culture collection center, deposit number: CCTCC No:
M2019050。
The separation of lactobacillus plantarum: it weighs 10g straw sample and is added in the 90ml distilled water of sterilizing, be coated with after gradient dilution
On MRS solid medium, 48h is cultivated according to form and color and grows bacterium colony on picking MRS, by producing sour, acidproof, temperature
The experiment sievings such as test produce that the acidproof resistance of acid is strong, inhibits 1 strains of lactic acid bacteria of pathogen, 24 hours lactic acid producing amount 1.51mg/
L, the inhibition zone to E.coli K88, staphylococcus aureus are respectively 21.4mm, 20.8mm.Bacterium colony is creamy white, and edge is whole
Together；Cell microscopic morphology is rod-short, no gemma, Gram-positive；Catalase is negative, oxidase negative；With relatively strong
Growth and product acid activity initial ph value be 3.0-8.0 culture medium on can grow, faint growth is presented under the conditions of 10 DEG C,
Upgrowth situation is good under the conditions of 15 DEG C -45 DEG C, is accredited as lactobacillus plantarum through 16rRNA gene order, is preserved in Chinese allusion quotation
Type culture collection, deposit number: CCTCC No:M2016281.
The preparation of 2 complex micro organism fungicide of embodiment
Amplification of the configuration of the composite bacteria agent from slant strains to finished product will pass through following three steps
1, level-one spreads cultivation: the strain of each purified culture (bacillus amyloliquefaciens, bacillus megaterium, long shoot trichoderma,
Lactobacillus plantarum) it after taking-up, is inoculated into four seed culture mediums prepared and cultivates respectively respectively, often from the inclined-plane of preservation
Culture medium 200ml is filled in the bottle of a 1L, lactobacillus plantarum cultivates 48h in 37 DEG C of constant incubator, bacillus amyloliquefaciens,
Bacillus megaterium 35 DEG C of 150r/min shaken cultivation 48h, long shoot trichodermas 30 DEG C of 120r/min oscillations on shaking table on shaking table
Cultivate 96h.
In level-one spreads cultivation, the slant medium and seed culture medium of various bacterium are well known basal medium.Wherein
Bacillus amyloliquefaciens, bacillus megaterium seed culture medium --- nutrient broth medium (culture bacterium), long shoot wood
It is mould --- PDA culture medium (culture mould), lactobacillus plantarum --- MRS culture medium (culture lactic acid bacteria).
2, second level spreads cultivation: period detects each bacteria culture fluid viable count respectively, to each bacterium viable count >=2 × 109After/ml, take
Four kinds of bacterium are pressed percent by volume bacillus amyloliquefaciens 25%, bacillus megaterium 30%, long shoot trichoderma by seed liquor out
25%, composite bacteria liquid is made in lactobacillus plantarum 20%, is then composite bacteria liquid 10% and solid medium 90% according to weight ratio
Ratio composite bacteria liquid is inoculated in solid medium, mixing is collectively disposed under 35 DEG C of condition of culture and cultivates 72h.
Second level spread cultivation in solid culture based formulas (weight percent): soybean residue (aqueous 80%) 64%, wheat bran 22%,
Corn flour 10%, glucose 3.5%, NaCl 0.4%, KH2PO40.1%, pH6.4-7.2.
3) expect drying admittedly: drying can be made into complex micro organism fungicide under the conditions of 50 DEG C.Microbial inoculum bacteria containing amount is 4 × 109-
8×109Between cfu/g
Embodiment 3: use compound microbial culture starter of the invention in the bed body effect test of cage bird fermentation bed
In December, -2017 in October, 2017 has carried out application experiment of the product of the present invention in bed surface matrix in certain chicken house.
Cage bird ferments bed 40cm or so mushroom bran, and surface spreads sawdust/rice husk 10cm or so, and sawdust and rice husk weight ratio are 1:1, with turning over
The operation stirring of rake machine, padding is mixed, padding is with a thickness of 50cm.Every square metre of fermentation bed of test group with 50 grams of fermentation bed bacteria,
It is sprinkled into method be first by fermentation bed bacteria, wheat bran after mixing, be spread on fermentation bed surface, with machine for turning up the soil and harrowing run stir；Hair
The weight ratio of ferment bed microbial inoculum, wheat bran are as follows: fermentation bed bacteria 50g, wheat bran 1kg.Every square metre of control group only spreads 1kg wheat bran.
Fermenting bed padding is taken in 15d, 30d, 45d, 60d respectively, monitors microorganism and the nutrient content of padding.Total bacterium is used
37 DEG C of constant temperature incubations of nutrient agar (NA) culture medium are for 24 hours；The dilution of bacillus uses nutrient agar after 80 DEG C of water-bath 10min
(NA) 37 DEG C of constant temperature incubations of culture medium are for 24 hours；Fungi 28 DEG C of culture 5d of potato dextrose agar；Staphylococcus aureus
Bacterium, Escherichia coli use 37 DEG C of mannitol sodium chloride agar (MSA), eosin methylene blue agar culture medium (EMB) constant temperature incubations respectively
48h.Nitrobacteria Stefansson culture solution MPN method, denitrifying bacteria use sodium citrate potassium nitrate culture medium MPN method.Ammonia
State nitrogen is using potassium chloride extraction-indophenol blue colorimetry measurement.Organic matter is measured using potassium dichromate oxidation.Padding humus
And its component uses sodium pyrophosphate-sodium hydroxide solution extraction method, potassium dichromate method measurement.As a result as shown in table 1, table 2, table 3.
By table 1,2 it is found that adding microbial bacterial agent of the invention significantly improves bacillus distributed quantity, golden yellow grape is significantly reduced
Coccus distributed quantity significantly improves the nitrobacteria (P < 0.05) in nitrogen metabolism microorganism, reduces nitrogen loss.By table 3
It is found that adding microbial bacterial agent of the invention can significantly reduce the water content and ammonia nitrogen content of padding, to significantly improve fowl
Environment is given up, in terms of padding Fertilizer Transformed, addition microbial inoculum significantly improves the content of humic acid and total humus, improves padding humic
Rate forms high-quality organic fertilizer.
The padding microorganism (LogCFU/g) of 1 cage bird fermentation bed of table
The nitrogen metabolism microorganism (LogCFU/g) of 2 cage bird fermenting bed padding of table
The nutrient content of 3 cage bird fermenting bed padding of table
For the microbiota for further studying cage bird fermentation bed, cage is studied using molecular microbiology method system
The microorganism group of chicken padding is at the microbial DNA of extraction 60d padding sample has carried out the amplification in the area 16SrDNAV3, used
HiSeq platform sequencing analysis.As a result as shown in table 4 and Fig. 1.Shannon diversity index, Simpson's indexes Index are the micro- lifes of reflection
The multifarious important indicator of object.Shannon diversity index is higher, and Simpson's indexes Index is lower, shows that the diversity of microorganism is got over
It is high.As shown in Table 4, addition fermentation bed bacteria significantly improves the diversity of chicken padding microorganism, significantly promotes microorganism pair
The ira situ degradation of chicken manure urine.LEfSe analysis is carried out to different groups of padding microbial DNAs, finds the rich of two group dominant microfloras
Degree variation.As a result as shown in Figure 1, addition chicken fermentation bed bacteria significantly improves bacillus in padding (Bacillus), sea
The flora abundance of foreign Bacillus (Oceanobacillus), significantly reduces the flora of popular Bacillus (Vulgatibacter)
Abundance.Bacillus, ocean Bacillus are the critical bacterial populations for participating in excrement conversion.Test result shows to add fermentation bed bacterium
Agent, bacillus in microbial inoculum can in chicken manure padding fast-growth, form dominant microflora, promote the ira situ degradation of chicken manure urine.
The Analysis of Microbial Diversity of the different padding samples of table 4
Claims (10)
1. a kind of microbial composite for being used to prepare cage bird fermenting bed padding, it is characterised in that by the micro- of following volumes proportion
Bio-fermented liquid composition: bacillus amyloliquefaciens fermentation liquid 20-30%, bacillus megaterium fermentation liquid 25-35%, long shoot trichoderma
Fermentation liquid 20-30%, lactobacillus plantarum fermentation liquid 15-25%, total viable count contained by microbial inoculum is 4 × 109-8×109Cfu/g it
Between.
2. microbial composite according to claim 1, it is characterised in that the microbial fermentation solution matched by following volumes
Composition: bacillus amyloliquefaciens 25%, bacillus megaterium 30%, long shoot trichoderma 25%, lactobacillus plantarum 20%, contained by microbial inoculum
Total viable count 4 × 109-8×109Between cfu/g.
3. a kind of complex micro organism fungicide for being used to prepare cage bird fermenting bed padding, it is characterised in that the complex microorganism
Microbial inoculum is that microbial composite of any of claims 1 or 2 is inoculated in solid medium, and mixing is collectively disposed at 30-35 DEG C
72-96h is cultivated under condition of culture, drying is made.
4. complex micro organism fungicide according to claim 3, it is characterised in that by microorganism of any of claims 1 or 2
Microbial composite is inoculated in solid medium by composition and solid medium according to the ratio that weight ratio is 1:9, is mixed
It is collectively disposed under 30-35 DEG C of condition of culture and cultivates 72-96h；Then drying can be made into composite microbial bacteria under the conditions of 50 DEG C
Agent；The solid culture based formulas are as follows: soybean residue 64%, wheat bran 22%, corn flour 10%, glucose 3.5%,
NaCl0.4%, KH2PO40.1%, pH6.4-7.2.
5. the preparation method of complex micro organism fungicide as claimed in claim 3, it is characterised in that including by claims 1 or 2 institute
The microbial composite stated is inoculated in solid medium, and mixing, which is collectively disposed under 30-35 DEG C of condition of culture, cultivates 72-96h, is dried
It is drying to obtain the complex micro organism fungicide.
6. preparation method according to claim 5, it is characterised in that the solid culture based formulas are as follows: soybean residue
64%, wheat bran 22%, corn flour 10%, glucose 3.5%, NaCl0.4%, KH2PO40.1%, pH6.4-7.2；Claim
The weight ratio of microbial composite described in 1 or 2 and solid medium is 1:9.
7. complex micro organism fungicide described in claim 3 or 4 is preparing the application in cage bird fermenting bed padding.
8. a kind of cage bird fermenting bed padding, it is characterised in that include complex micro organism fungicide described in claim 3 or 4.
9. cage bird fermenting bed padding according to claim 8, it is characterised in that the cage bird fermenting bed padding is also
Including mushroom bran, sawdust, rice husk and wheat bran.
10. cage bird fermenting bed padding according to claim 8, it is characterised in that cage bird fermentation 35~40cm of bed bacterium
Chaff, surface spread the weight ratios such as sawdust and rice husk 8~15cm of mixture, are run and are stirred with machine for turning up the soil and harrowing, by mushroom bran and sawdust and rice husk
Mixture mixing；After mixing by complex micro organism fungicide, wheat bran described in claim 3 or 4, it is spread on fermentation bed table
Face, then run and stirred with machine for turning up the soil and harrowing.Wherein；The weight ratio of complex micro organism fungicide described in claim 3 or 4, wheat bran are as follows: 1:
15~22；Every square of padding spreads complex micro organism fungicide 50g described in claim 3 or 4.
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