CN101974434A - Multi-bacterium microbial compound preparations for aquaculture and preparation method thereof - Google Patents
Multi-bacterium microbial compound preparations for aquaculture and preparation method thereof Download PDFInfo
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- CN101974434A CN101974434A CN2010105403087A CN201010540308A CN101974434A CN 101974434 A CN101974434 A CN 101974434A CN 2010105403087 A CN2010105403087 A CN 2010105403087A CN 201010540308 A CN201010540308 A CN 201010540308A CN 101974434 A CN101974434 A CN 101974434A
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- 238000002360 preparation method Methods 0.000 title claims abstract description 69
- 238000009360 aquaculture Methods 0.000 title claims abstract description 38
- 230000000813 microbial effect Effects 0.000 title claims abstract description 28
- 150000001875 compounds Chemical class 0.000 title claims abstract description 24
- 244000144974 aquaculture Species 0.000 title abstract description 19
- 238000011218 seed culture Methods 0.000 claims abstract description 111
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 59
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 56
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 52
- 230000001580 bacterial effect Effects 0.000 claims abstract description 30
- 241001148165 Rhodoplanes elegans Species 0.000 claims abstract description 21
- 235000014897 Streptococcus lactis Nutrition 0.000 claims abstract description 21
- 239000012531 culture fluid Substances 0.000 claims description 107
- 241000894006 Bacteria Species 0.000 claims description 58
- 235000015097 nutrients Nutrition 0.000 claims description 47
- 239000007788 liquid Substances 0.000 claims description 44
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 31
- 241000186660 Lactobacillus Species 0.000 claims description 30
- 230000000243 photosynthetic effect Effects 0.000 claims description 30
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 29
- 229940039696 lactobacillus Drugs 0.000 claims description 27
- 241000235648 Pichia Species 0.000 claims description 20
- 244000057717 Streptococcus lactis Species 0.000 claims description 20
- 239000012153 distilled water Substances 0.000 claims description 17
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 12
- 239000001301 oxygen Substances 0.000 claims description 12
- 229910052760 oxygen Inorganic materials 0.000 claims description 12
- 239000001888 Peptone Substances 0.000 claims description 9
- 108010080698 Peptones Proteins 0.000 claims description 9
- 235000019319 peptone Nutrition 0.000 claims description 9
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 6
- 235000012907 honey Nutrition 0.000 claims description 5
- 239000002054 inoculum Substances 0.000 claims description 5
- 241001037822 Bacillus bacterium Species 0.000 claims description 3
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 3
- 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 3
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 3
- 229910019142 PO4 Inorganic materials 0.000 claims description 3
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims description 3
- 229920002472 Starch Polymers 0.000 claims description 3
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 3
- 240000008042 Zea mays Species 0.000 claims description 3
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 3
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 3
- 239000001110 calcium chloride Substances 0.000 claims description 3
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 3
- 239000004202 carbamide Substances 0.000 claims description 3
- 235000005822 corn Nutrition 0.000 claims description 3
- 239000011790 ferrous sulphate Substances 0.000 claims description 3
- 235000003891 ferrous sulphate Nutrition 0.000 claims description 3
- 239000008103 glucose Substances 0.000 claims description 3
- 238000005286 illumination Methods 0.000 claims description 3
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 3
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 3
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 claims description 3
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 claims description 3
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 3
- 239000010452 phosphate Substances 0.000 claims description 3
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 3
- 229920000053 polysorbate 80 Polymers 0.000 claims description 3
- 239000011591 potassium Substances 0.000 claims description 3
- 229910052700 potassium Inorganic materials 0.000 claims description 3
- 235000019698 starch Nutrition 0.000 claims description 3
- 239000008107 starch Substances 0.000 claims description 3
- 235000015193 tomato juice Nutrition 0.000 claims description 3
- 241001465754 Metazoa Species 0.000 abstract description 16
- 201000010099 disease Diseases 0.000 abstract description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 3
- 241000521553 Pichia fermentans Species 0.000 abstract 2
- 241000186000 Bifidobacterium Species 0.000 abstract 1
- 241000194035 Lactococcus lactis Species 0.000 abstract 1
- 238000012258 culturing Methods 0.000 abstract 1
- 238000000855 fermentation Methods 0.000 abstract 1
- 230000004151 fermentation Effects 0.000 abstract 1
- 239000001963 growth medium Substances 0.000 abstract 1
- 241000238557 Decapoda Species 0.000 description 11
- 238000012360 testing method Methods 0.000 description 9
- 238000000034 method Methods 0.000 description 7
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 6
- 239000002253 acid Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 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 3
- 241000196324 Embryophyta Species 0.000 description 3
- 235000001014 amino acid Nutrition 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 3
- FPVGTPBMTFTMRT-UHFFFAOYSA-L disodium;2-amino-5-[(4-sulfonatophenyl)diazenyl]benzenesulfonate Chemical class [Na+].[Na+].C1=C(S([O-])(=O)=O)C(N)=CC=C1N=NC1=CC=C(S([O-])(=O)=O)C=C1 FPVGTPBMTFTMRT-UHFFFAOYSA-L 0.000 description 3
- 235000014655 lactic acid Nutrition 0.000 description 3
- 230000000050 nutritive effect Effects 0.000 description 3
- 239000004382 Amylase Substances 0.000 description 2
- 102000013142 Amylases Human genes 0.000 description 2
- 108010065511 Amylases Proteins 0.000 description 2
- 241001609213 Carassius carassius Species 0.000 description 2
- 241000195493 Cryptophyta Species 0.000 description 2
- 241000252230 Ctenopharyngodon idella Species 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 241000252234 Hypophthalmichthys nobilis Species 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 235000019418 amylase Nutrition 0.000 description 2
- 244000052616 bacterial pathogen Species 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- 230000003628 erosive effect Effects 0.000 description 2
- 238000012851 eutrophication Methods 0.000 description 2
- 239000013505 freshwater Substances 0.000 description 2
- 244000000010 microbial pathogen Species 0.000 description 2
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 1
- 102000004882 Lipase Human genes 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- UCKMPCXJQFINFW-UHFFFAOYSA-N Sulphide Chemical compound [S-2] UCKMPCXJQFINFW-UHFFFAOYSA-N 0.000 description 1
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 1
- 239000004473 Threonine Substances 0.000 description 1
- -1 Xie Ansuan Chemical compound 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 230000007969 cellular immunity Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000000378 dietary effect Effects 0.000 description 1
- 230000005059 dormancy Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000004459 forage Substances 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 244000005709 gut microbiome Species 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
- Y02A40/818—Alternative feeds for fish, e.g. in aquacultures
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to multi-bacterium microbial compound preparations for aquaculture and a preparation method thereof. The multi-bacterium microbial compound preparations are characterized by comprising the following steps of: preparing seed culture solution by performing single strain culture on rhodoplanes elegans, pichia fermentans lodder, bacillus natto swamura, bacillus subtills, lactococcus lactis and bifidobacterium sp respectively; inoculating the seed culture solution onto the same culture medium; and then culturing the seed culture solution by a fermentation tank, and naturally reducing a pH value of the seed culture solution to 3.0 to 3.5 to obtain the multi-bacterium microbial compound preparations, wherein the bacterial content of the pichia fermentans lodder in the preparations is more than or equal to 0.1*10<9>cfu/ml; the bacterial content of the other strains is more than or equal to 5.0*10<9>cfu/ml. The multi-bacterium microbial compound preparations for aquaculture can improve the ecological environment of aquatic animals, prevents and treats diseases, reduces the emergency condition of the animals, improves the quality of the aquatic animals, and shortens a listed period.
Description
Technical field
The invention belongs to aquaculture field, particularly a kind of many bacterial classifications of used for aquiculture microbial compound preparation and preparation method thereof.
Background technology
A large amount of appearance along with the mass-producing culture fishery; cultivation density constantly increases; the movement in the aquaculture water and the residue of bait constantly increase; pathogenic microorganism is bred in a large number; the aggravation of aquaculture water eutrophication degree; ecotope suffers serious destruction; this causes aquatic animal to occur all kinds of diseases and emergency rating on a large scale and frequently; meanwhile the life-time service of microbiotic and sterilant and abuse also further caused the residual and pathogenic microorganism resistance of various medicines in the aquaculture water and chemical sproof raising with, thereby further destroyed the ecotope of aquaculture water.
In recent years, microbial preparation is existing the application in aquaculture, but also exists many problems.The most of microbe preparation is only brought into play effect at aquaculture water, but to not directly effect of aquatic animal itself.
Summary of the invention
Technical problem to be solved by this invention provides a kind of many bacterial classifications of used for aquiculture microbial compound preparation and preparation method thereof.This many bacterial classifications of used for aquiculture microbial compound preparation can improve the ecotope of aquatic animal simultaneously, and disease preventing and treating reduces the animal emergency rating, improves the aquatic animal quality, shortens the listing cycle.
For solving the problems of the technologies described above, the technical solution used in the present invention is:
A kind of many bacterial classifications of used for aquiculture microbial compound preparation is characterized in that: it be by photosynthetic bacterium (
Rhodoplanes elegans) ACCC 10649, yeast (
Pichia fermentansLodder) ACCC 20319
T, Bacillus natto (
Bacillus natto Swamura) ACCC 10614, subtilis (
Bacillus subtillsSubsp.
SpizieniiNakamura et al) ACCC 10242
T, lactobacillus (
Lactococcus lactis) ACCC 11092 and bifidus bacillus (
BifidobacteriumSp) ACCC 11054 carries out the seed culture fluid that single spawn culture obtains respectively and is inoculated in the same substratum, carries out fermentor cultivation then, and the pH value for the treatment of nutrient solution is reduced to 3.0～3.5 naturally and obtained, photosynthetic bacterium in the said preparation (
Rhodoplanes elegans) bacterium number 〉=5.0 * 10 of ACCC 10649
9Cfu/ml, yeast (
Pichia fermentansLodder) ACCC 20319
TBacterium number 〉=0.1 * 10
9Cfu/ml, Bacillus natto (
Bacillus natto Swamura) bacterium number 〉=5.0 * 10 of ACCC 10614
9Cfu/ml, subtilis (
Bacillus subtillsSubsp.
SpizieniiNakamura et al) ACCC 10242
TBacterium number 〉=5.0 * 10
9Cfu/ml, lactobacillus (
Lactococcus lactis) bacterium number 〉=5.0 * 10 of ACCC 11092
9Cfu/ml, bifidus bacillus (
BifidobacteriumSp) bacterium number 〉=5.0 * 10 of ACCC 11054
9Cfu/ml.
Press such scheme, the weight percentage of the seed culture fluid of described each bacterial classification is: the seed culture fluid 25%～35% of photosynthetic bacterium, saccharomycetic seed culture fluid 5%～15%, the seed culture fluid 15%～25% of Bacillus natto, the seed culture fluid 15%～25% of subtilis, the seed culture fluid 5%～15% of lactobacillus, the seed culture fluid 5%～15% of bifidus bacillus.
A kind of preparation method of many bacterial classifications of used for aquiculture microbial compound preparation is characterized in that: comprise the steps:
(1) preparation of seed culture fluid:
Photosynthetic bacterium (
Rhodoplanes elegans) preparation of seed culture fluid of ACCC 10649: with photosynthetic bacterium (
Rhodoplanes elegans) ACCC 10649 is inoculated in the liquid nutrient medium under aseptic condition, described liquid nutrient medium is: the content of each component in 1.0L distilled water is: yeast powder 3.0g, peptone 3.0g, magnesium sulfate heptahydrate 0.05g, calcium chloride 0.3g, the pH value transfers between 6.08～7.0, be positioned in the light incubator, 28 ℃ of-30 ℃ of conventional illumination cultivation 6～7 days obtain photosynthetic bacterium bacterium number 〉=5.0 * 10
9The seed culture fluid of cfu/ml;
Yeast (
Pichia fermentansLodder) ACCC 20319
TThe preparation of seed culture fluid: with yeast (
Pichia fermentansLodder) ACCC 20319
TUnder aseptic condition, be inoculated in the liquid nutrient medium, described liquid nutrient medium is: the content of each component in 1.0L distilled water is: anhydrous magnesium sulfate 1.0g, potassium primary phosphate 2.0g, urea 3.0g, corn steep liquor 1.0g, manganous sulfate 0.02g, ferrous sulfate 0.2g, the pH value transfers to 5.5, cultivated 4～5 days, and obtained yeast bacterium number 〉=0.1 * 10 for 30 ℃-31 ℃
9The seed culture fluid of cfu/ml;
Bacillus natto (
Bacillus natto Swamura) seed culture fluid of ACCC10614 and subtilis (
Bacillus subtillsSubsp.
SpizieniiNakamura et al) ACCC 10242
TThe preparation of seed culture fluid: with Bacillus natto (
Bacillus natto Swamura) ACCC 10614 and subtilis (
Bacillus subtillsSubsp.
SpizieniiNakamura et al) ACCC 10242
TUnder aseptic condition, be inoculated in the liquid nutrient medium respectively, described liquid nutrient medium is: the content of each component in 1.0L distilled water is: peptone 10.0g, extractum carnis 3.0g, sodium-chlor 5.0g, the pH value transfers to 7.0,28 ℃～32 ℃, single spawn culture 7～8 days obtains Bacillus natto bacterium number 〉=5.0 * 10 respectively
9The Bacillus natto seed culture fluid of cfu/ml and bacterial count of bacillus subtilis 〉=5.0 * 10
9The subtilis seed culture fluid of cfu/ml;
Lactobacillus (
Lactococcus lactis) seed culture fluid of ACCC 11092 and bifidus bacillus (
BifidobacteriumSp) preparation of the seed culture fluid of ACCC 11054: with lactobacillus (
Lactococcus lactis) ACCC 11092 and bifidus bacillus (
BifidobacteriumSp) ACCC 11054 is inoculated in the liquid nutrient medium under aseptic condition respectively, described liquid nutrient medium is: the content of each component in 1.0L distilled water is: tomato juice 200.0ml, Zulkovsky starch 0.5g, peptone 15.0g, sodium-chlor 5.0g, yeast powder 6.0g, tween 80 1.0g, glucose 20.0g, the pH value transfers to 7.2,30 ℃～35 ℃ single spawn culture 2～3 days obtain lactobacillus bacterium number 〉=5.0 * 10 respectively
9The lactobacillus seed nutrient solution of cfu/ml and bifidus bacillus bacterium number 〉=5.0 * 10
9The bifidus bacillus seed nutrient solution of cfu/ml;
(2) preparation of many bacterial classifications of used for aquiculture microbial compound preparation:
Respectively the photosynthetic bacterium that step (1) is prepared (
Rhodoplanes elegans) ACCC 10649, yeast (
Pichia fermentansLodder) ACCC 20319
T, Bacillus natto (
Bacillus natto Swamura) ACCC10614, subtilis (
Bacillus subtillsSubsp.
SpizieniiNakamura et al) ACCC 10242
T, lactobacillus (
Lactococcus lactis) ACCC 11092 and bifidus bacillus (
BifidobacteriumSp) seed culture fluid of ACCC 11054 by its weight percentage is: the seed culture fluid 25%～35% of photosynthetic bacterium, saccharomycetic seed culture fluid 5%～15%, the seed culture fluid 15%～25% of Bacillus natto, the seed culture fluid 15%～25% of subtilis, the seed culture fluid 5%～15% of lactobacillus, the seed culture fluid 5%～15% of bifidus bacillus, under aseptic condition, be inoculated in the same liquid nutrient medium respectively, inoculum size is 1 ﹪～2 ﹪ of liquid nutrient medium weight, the content of each component in 1.0L distilled water is in the described liquid nutrient medium: brown sugar 50.0g, honey 10.0g, at oxygen dissolved is that 5 ﹪ are that the volume ratio of the volume of oxygen in the fermentor tank and fermented liquid is under 5% the condition, 28 ℃～32 ℃ closeds leave standstill cultivated 7～15 days, made the pH value drop to 3.0-3.5 and formed.
Utilize the characteristic of different strain can correspondingly solve the problem that occurs in the aquaculture, but different strain microorganism compound tense exist to reject, seize Tan Yuan ∕ nitrogenous source and mutual erosive phenomenon mutually between each bacterial classification.The present invention is fermented the bacterial classification of different qualities respectively, obtain seed culture fluid, be inoculated in mixed culture in the same substratum then by a certain percentage respectively, naturally being reduced to 3.0～3.5 to the pH value forms, under this pH condition, each bacterial classification is in extreme environment, forces each bacterial classification to enter dormant state, thereby the mutual rejection that can avoid existing between the different strain, seizes Tan ∕ nitrogenous source and erosive phenomenon mutually.In use, with the used for aquiculture microbial compound preparation of the present invention preparation put into culture the waters after because the change of environment can make bacterial classification breaking dormancy state, thereby each bacterial classification is played one's part to the full, solve the problem that occurs in the actual aquaculture.
The function of the bacterial classification that the present invention is selected is as follows:
(1) photosynthetic bacterium (
Rhodoplanes elegans) ACCC 10649: consume unnecessary nutritive substance in the water body, produce a large amount of useful nutritive substances, reduce objectionable impuritiess such as ammonia nitrogen, sulfide aquatic animal.
(2) yeast (
Pichia fermentansLodder) ACCC 20319
T: produce the essential multiple amino acids of animal in a large number.
(3) subtilis (
Bacillus subtillsSubsp.
SpizieniiNakamura et al) ACCC 10242
T: produce proteolytic enzyme, amylase and subtilyne.
(4) lactobacillus (
Lactococcus lactis) ACCC 11092: can decompose sugar and produce acid, suppress the breeding of pathogenic bacterium and spoilage organism, improve the absorption of the bait of animal.
(5) bifidus bacillus (
BifidobacteriumSp) ACCC 11054: safeguard enteron aisle normal bacteria colony balance, suppress growth of pathogenic bacteria, synthesise vitamins, amino acid and raising body produce VITMAIN B1, B2, B6, B12 and L-Ala, Xie Ansuan, aspartic acid and Threonine etc. to the absorption of calcium ion in enteron aisle.
(6) Bacillus natto (
Bacillus natto Swamura) ACCC10614: regulate intestinal microflora, the enhancing animal cell immunity is put should, and can generate multiple protein enzyme (particularly Sumizyme MP), saccharifying enzyme, lipase, amylase, some complicated carbohydrate of degrading plant forage kind, thereby the transformation efficiency of raising feed.
Beneficial effect of the present invention:
1. the recovery of micro-ecological environment in the promotion gi tract.Lactobacillus in this preparation (
Lactococcus lactis) ACCC 11092, bifidus bacillus (
BifidobacteriumSp) ACCC 11054, Bacillus natto (
Bacillus natto Swamura) ACCC 10614 can grow rapidly in the enteron aisle of aquatic animal surely, sets up the population advantage of probiotics, promotes the dietetic alimentation of aquatic animal, produces multiple aquatic animal grow necessary amino acid, VITAMIN and organic acid, and bifidus bacillus (
BifidobacteriumSp) ACCC 11054, Bacillus natto (
Bacillus natto Swamura) ACCC10614 enhancing body's immunological function significantly, effectively prevent digestive tract infection, reduce emergency rating;
2. purifying aquatic water.Photosynthetic bacterium in this preparation (
Rhodoplanes elegans) ACCC 10649, yeast (
Pichia fermentansLodder) ACCC 20319
TAnd subtilis (
Bacillus subtillsSubsp.
SpizieniiNakamura et al) ACCC 10242
TCan in time decompose residual bait in animal excrements, metabolite and the water body that in the aquaculture process, produces, obviously reduce the eutrophy degree of aquaculture water, improve the oxygen dissolved of water body, produce multiple proteins simultaneously, for aquatic animal provides the fine natural bait, also for planktonic organism provides growth needed nutritive substance, thereby further set up and improve the micro-ecological environment of aquaculture water.
Embodiment
The present invention is described in further detail with specific embodiment below, but the present invention not only is confined to following examples.
Embodiment 1
The preparation method of used for aquiculture microbial preparation comprises the steps:
(1) preparation of seed culture fluid:
Photosynthetic bacterium (
Rhodoplanes elegans) the seed culture fluid preparation of ACCC 10649: with photosynthetic bacterium (
Rhodoplanes elegans) ACCC 10649 is under aseptic condition, be inoculated in the liquid nutrient medium, the content of each component in 1.0L distilled water is in the described liquid nutrient medium: yeast powder 3.0g, peptone 3.0g, magnesium sulfate heptahydrate 0.05 gram, calcium chloride 0.3g, the pH value transfers between 6.08～7.0, be positioned in the conventional light incubator, 28 ℃～30 ℃ conventional illumination cultivation 6～7 days obtain photosynthetic bacterium bacterium number 〉=5.0 * 10
9The seed culture fluid of cfu/ml.
Yeast (
Pichia fermentansLodder) ACCC 20319
TThe preparation of seed culture fluid: with yeast (
Pichia fermentansLodder) ACCC 20319
TUnder aseptic condition, be inoculated in the liquid nutrient medium, the content of each component in 1.0L distilled water is in the described liquid nutrient medium: anhydrous magnesium sulfate 1.0g, potassium primary phosphate 2.0g, urea 3.0g, corn steep liquor 1.0g, manganous sulfate 0.02g, ferrous sulfate 0.2g, the pH value transfers to 5.5, cultivated 4～5 days, and obtained yeast bacterium number 〉=0.1 * 10 for 30 ℃-31 ℃
9The seed culture fluid of cfu/ml.
Bacillus natto (
Bacillus natto Swamura) ACCC 10614 seed culture fluids and subtilis (
Bacillus subtillsSubsp.
SpizieniiNakamura et al) ACCC 10242
TThe preparation of seed culture fluid: with Bacillus natto (
Bacillus natto Swamura) ACCC 10614 and subtilis (
Bacillus subtillsSubsp.
SpizieniiNakamura et al) ACCC 10242
TUnder aseptic condition, be inoculated in the liquid nutrient medium respectively, described liquid nutrient medium is: the content of each component in 1.0L distilled water is: peptone 10.0g, extractum carnis 3.0g, sodium-chlor 5.0g, the pH value transfers to 7.0,28 ℃～32 ℃ single spawn culture 7～8 days obtain Bacillus natto bacterium number 〉=5.0 * 10 respectively
9The Bacillus natto seed culture fluid of cfu/ml and bacterial count of bacillus subtilis 〉=5.0 * 10
9The subtilis seed culture fluid of cfu/ml.
Lactobacillus (
Lactococcus lactis) ACCC 11092 seed culture fluids and bifidus bacillus (
BifidobacteriumSp) preparation of ACCC 11054 seed culture fluids: with lactobacillus (
Lactococcus lactis) ACCC 11092 and bifidus bacillus (
BifidobacteriumSp) ACCC 11054 is inoculated in the same liquid nutrient medium under aseptic condition respectively, the content of each component in 1.0L distilled water is in the described liquid nutrient medium: tomato juice 200.0mL, Zulkovsky starch 0.5g, peptone 15.0g, sodium-chlor 5.0g, yeast powder 6.0g, tween 80 1.0g, glucose 20.0g, the pH value transfers to 7.2,30 ℃～35 ℃ single spawn culture 2～3 days obtain lactobacillus bacterium number 〉=5.0 * 10 respectively
9The lactobacillus seed nutrient solution of cfu/ml and bifidus bacillus bacterium number 〉=5.0 * 10
9The bifidus bacillus seed nutrient solution of cfu/ml.
(2) preparation of used for aquiculture microbial preparation:
Respectively the seed culture fluid of each bacterial classification that step (1) is prepared according to photosynthetic bacterium (
Rhodoplanes elegans) ACCC 10649 seed culture fluid, yeast (
Pichia fermentansLodder) ACCC 20319
TSeed culture fluid, Bacillus natto (
Bacillus natto Swamura) ACCC10614 seed culture fluid, subtilis (
Bacillus subtillsSubsp.
SpizieniiNakamura et al) ACCC 10242
TSeed culture fluid, milk-acid bacteria (
Lactococcus lactis) seed culture fluid of ACCC 11092 and bifidus bacillus (
BifidobacteriumSp) weight percentage of the seed culture fluid of ACCC 11054 is: the seed culture fluid 25% of photosynthetic bacterium, saccharomycetic seed culture fluid 5%, the seed culture fluid 15% of Bacillus natto, the seed culture fluid 25% of subtilis, the seed culture fluid 15% of lactobacillus, the seed culture fluid 15% of bifidus bacillus, under aseptic condition, be inoculated in the same liquid nutrient medium respectively, inoculum size is 1 ﹪～2 ﹪ of liquid nutrient medium weight, the content of each component in 1.0L distilled water is in the described liquid nutrient medium: brown sugar 50.0g, honey 10.0g, oxygen dissolved be 5 ﹪ be in the fermentor tank oxygen volume and fermentating liquid volume than being under 5% the condition, 28 ℃～32 ℃ closeds leave standstill cultivated 7～15 days, made the pH value drop to 3.0-3.5 naturally and formed.
After testing, this used for aquiculture microbial compound preparation is acid, and golden yellow color, pH value are 3.0-3.5, and viable bacteria is respectively in its preparation: photosynthetic bacterium (
Rhodoplanes elegans) bacterium number 〉=5.0 * 10 of ACCC 10649
9Cfu/ml, yeast (
Pichia fermentansLodder) ACCC 20319
TBacterium number 〉=〉=0.1 * 10
9Cfu/ml, Bacillus natto (
Bacillus natto Swamura) bacterium number 〉=5.0 * 10 of ACCC 10614
9Cfu/ml, subtilis (
Bacillus subtillsSubsp.
SpizieniiNakamura et al) ACCC 10242
TBacterium number 〉=5.0 * 10
9Cfu/ml, lactobacillus (
Lactococcus lactis) bacterium number 〉=5.0 * 10 of ACCC 11092
9Cfu/ml, bifidus bacillus (
BifidobacteriumSp) bacterium number 〉=5.0 * 10 of ACCC 11054
9Cfu/ml.
Embodiment 2:
(1) preparation of seed culture fluid:
Adopt the method for present embodiment 1, make seed culture fluid;
(2) preparation of used for aquiculture microbial preparation:
Respectively the seed culture fluid of each bacterial classification that step (1) is prepared according to photosynthetic bacterium (
Rhodoplanes elegans) ACCC 10649 seed culture fluid, yeast (
Pichia fermentansLodder) ACCC 20319
TSeed culture fluid, Bacillus natto (
Bacillus natto Swamura) ACCC10614 seed culture fluid, subtilis (
Bacillus subtillsSubsp.
SpizieniiNakamura et al) ACCC 10242
TSeed culture fluid, milk-acid bacteria (
Lactococcus lactis) seed culture fluid of ACCC 11092 and bifidus bacillus (
BifidobacteriumSp) weight percentage of the seed culture fluid of ACCC 11054 is: the seed culture fluid 35% of photosynthetic bacterium, saccharomycetic seed culture fluid 15%, the seed culture fluid 25% of Bacillus natto, the seed culture fluid 15% of subtilis, the seed culture fluid 5% of lactobacillus, the seed culture fluid 5% of bifidus bacillus, under aseptic condition, be inoculated in the same liquid nutrient medium respectively, inoculum size is 1 ﹪～2 ﹪ of liquid nutrient medium weight, the content of each component in 1.0L distilled water is in the described liquid nutrient medium: brown sugar 50.0g, honey 10.0g, oxygen dissolved be 5 ﹪ be in the fermentor tank oxygen volume and fermentating liquid volume than being under 5% the condition, 28 ℃～32 ℃ closeds leave standstill cultivated 7～15 days, made the pH value drop to 3.0～3.5 naturally and formed.
After testing, this used for aquiculture microbial compound preparation is acid, and golden yellow color, pH value are 3.0-3.5, and the viable count of each bacterial classification is respectively in its preparation: photosynthetic bacterium (
Rhodoplanes elegans) bacterium number 〉=5.0 * 10 of ACCC 10649
9Cfu/ml, yeast (
Pichia fermentansLodder) ACCC 20319
TBacterium number 〉=0.1 * 10
9Cfu/ml, Bacillus natto (
Bacillus natto Swamura) bacterium number 〉=5.0 * 10 of ACCC 10614
9Cfu/ml, subtilis (
Bacillus subtillsSubsp.
SpizieniiNakamura et al) ACCC 10242
TBacterium number 〉=5.0 * 10
9Cfu/ml, lactobacillus (
Lactococcus lactis) bacterium number 〉=5.0 * 10 of ACCC 11092
9Cfu/ml, bifidus bacillus (
BifidobacteriumSp) bacterium number 〉=5.0 * 10 of ACCC 11054
9Cfu/ml.
Embodiment 3:
(1) preparation of seed culture fluid:
Adopt the method for present embodiment 1, make seed culture fluid;
(2) preparation of used for aquiculture microbial preparation:
Respectively the seed culture fluid of each bacterial classification that step (1) is prepared according to photosynthetic bacterium (
Rhodoplanes elegans) ACCC 10649 seed culture fluid, yeast (
Pichia fermentansLodder) ACCC 20319
TSeed culture fluid, Bacillus natto (
Bacillus natto Swamura) ACCC10614 seed culture fluid, subtilis (
Bacillus subtillsSubsp.
SpizieniiNakamura et al) ACCC 10242
TSeed culture fluid, milk-acid bacteria (
Lactococcus lactis) seed culture fluid of ACCC 11092 and bifidus bacillus (
BifidobacteriumSp) weight percentage of the seed culture fluid of ACCC 11054 is: the seed culture fluid 30% of photosynthetic bacterium, saccharomycetic seed culture fluid 10%, the seed culture fluid 20% of Bacillus natto, the seed culture fluid 20% of subtilis, the seed culture fluid 10% of lactobacillus, the seed culture fluid 10% of bifidus bacillus, under aseptic condition, be inoculated in the same liquid nutrient medium respectively, inoculum size is 1 ﹪～2 ﹪ of liquid nutrient medium weight, the content of each component in 1.0L distilled water is in the described liquid nutrient medium: brown sugar 50.0g, honey 10.0g, oxygen dissolved be 5 ﹪ be in the fermentor tank oxygen volume and fermentating liquid volume than being under 5% the condition, 28 ℃ of-32 ℃ of closeds leave standstill cultivated 7-15 days, made the pH value drop to 3.0～3.5 naturally and formed.
After testing, this used for aquiculture microbial compound preparation is acid, and golden yellow color, pH value are 3.0～3.5, and the viable count of each bacterial classification is respectively in its preparation: photosynthetic bacterium (
Rhodoplanes elegans) bacterium number 〉=5.0 * 10 of ACCC 10649
9Cfu/ml, yeast (
Pichia fermentansLodder) ACCC 20319
TBacterium number 〉=0.1 * 10
9Cfu/ml, Bacillus natto (
Bacillus natto Swamura) bacterium number 〉=5.0 * 10 of ACCC 10614
9Cfu/ml, subtilis (
Bacillus subtillsSubsp.
SpizieniiNakamura et al) ACCC 10242
TBacterium number 〉=5.0 * 10
9Cfu/ml, lactobacillus (
Lactococcus lactis) bacterium number 〉=5.0 * 10 of ACCC 11092
9Cfu/ml, bifidus bacillus (
BifidobacteriumSp) bacterium number 〉=5.0 * 10 of ACCC 11054
9Cfu/ml.
Application Example:
Adopt the aquaculture microbial compound preparation of the embodiment of the invention 1 preparation, in shrimp pond and fishpond, implement as follows respectively:
With the shrimp pond is example: test shrimp pond: 2 mu of areas, put in a suitable place to breed from 50,000/mu of numerous freshwater shrimp shrimp seedlings; Contrast shrimp pond: 2 mu of areas, put in a suitable place to breed from 50,000/mu of numerous freshwater shrimp shrimp seedlings.
Using method: in test shrimp pond, put in a suitable place to breed preceding 10 days in the shrimp seedling, adopt the Chi Shui method of splashing, according to 500ml aquaculture microbial compound preparation/mu, and it is diluted 200 times, per 10 days once, full pool spilling head three times.
Test-results: the dissolved oxygen comparison in test shrimp pond increases by 85% according to the pond; Ammonia-nitrogen content≤0.1mg/mL, comparison on average descends 65% according to the shrimp pond; The gross output comparison increases 25kg, amplification 15% according to the shrimp pond; Commodity rate reaches 90%, and specification is big, the price height, and contrast pond commodity rate is 54.5%, and specification is little, and price is low relatively.
With the fishpond is example:
The test fishpond: 8 mu of areas, put grass carp, crucian and silver carp fingerling in a suitable place to breed for every mu and amount to 90kg; The contrast fishpond: 8 mu of areas, put grass carp, crucian and silver carp fingerling in a suitable place to breed for every mu and amount to 90kg.
Using method: in the test fishpond, in fingerling stocking preceding 10 days, adopt the Chi Shui method of splashing, according to 500ml aquaculture microbial compound preparation/mu, and it is diluted 200 times, per 10 days are once, full pool spilling head three times.
Test-results: the per mu clean product comparison in test fishpond increases 100kg, amplification 25% according to the fishpond; Ammonia-nitrogen content≤0.1mg/mL, nitrite content≤0.02mg/mL, COD in water body reduces by 50%～60%, and body eutrophication effectively reduces; The plant plankton population number increases by 30%～40%, and part algae sum descends 60%～80%, has optimized the population structure of plant plankton in the water body, has suppressed the excessive multiplication of algae, sets up and the perfect micro-ecological environment of aquaculture water.
Claims (3)
1. many bacterial classifications of used for aquiculture microbial compound preparation is characterized in that: it be by photosynthetic bacterium (
Rhodoplanes elegans) ACCC 10649, yeast (
Pichia fermentansLodder) ACCC 20319
T, Bacillus natto (
Bacillus natto Swamura) ACCC 10614, subtilis (
Bacillus subtillsSubsp.
SpizieniiNakamura et al) ACCC 10242
T, lactobacillus (
Lactococcus lactis) ACCC 11092 and bifidus bacillus (
BifidobacteriumSp) ACCC 11054 carries out the seed culture fluid that single spawn culture obtains respectively and is inoculated in the same substratum, carries out fermentor cultivation then, and the pH value for the treatment of nutrient solution is reduced to 3.0～3.5 naturally and obtained, photosynthetic bacterium in the said preparation (
Rhodoplanes elegans) bacterium number 〉=5.0 * 10 of ACCC 10649
9Cfu/ml, yeast (
Pichia fermentansLodder) ACCC 20319
TBacterium number 〉=0.1 * 10
9Cfu/ml, Bacillus natto (
Bacillus natto Swamura) bacterium number 〉=5.0 * 10 of ACCC 10614
9Cfu/ml, subtilis (
Bacillus subtillsSubsp.
SpizieniiNakamura et al) ACCC 10242
TBacterium number 〉=5.0 * 10
9Cfu/ml, lactobacillus (
Lactococcus lactis) bacterium number 〉=5.0 * 10 of ACCC 11092
9Cfu/ml, bifidus bacillus (
BifidobacteriumSp) bacterium number 〉=5.0 * 10 of ACCC 11054
9Cfu/ml.
2. many bacterial classifications of used for aquiculture microbial compound preparation according to claim 1, it is characterized in that: the weight percentage of the seed culture fluid of described each bacterial classification is: the seed culture fluid 25%～35% of photosynthetic bacterium, saccharomycetic seed culture fluid 5%～15%, the seed culture fluid 15%～25% of Bacillus natto, the seed culture fluid 15%～25% of subtilis, the seed culture fluid 5%～15% of lactobacillus, the seed culture fluid 5%～15% of bifidus bacillus.
3. the preparation method of many bacterial classifications of used for aquiculture microbial compound preparation according to claim 1 is characterized in that: comprise the steps:
(1) preparation of seed culture fluid:
Photosynthetic bacterium (
Rhodoplanes elegans) preparation of seed culture fluid of ACCC 10649: with photosynthetic bacterium (
Rhodoplanes elegans) ACCC 10649 is inoculated in the liquid nutrient medium under aseptic condition, described liquid nutrient medium is: the content of each component in 1.0L distilled water is: yeast powder 3.0g, peptone 3.0g, magnesium sulfate heptahydrate 0.05g, calcium chloride 0.3g, the pH value transfers between 6.08～7.0, be positioned in the light incubator, 28 ℃～30 ℃ conventional illumination cultivation 6～7 days obtain photosynthetic bacterium bacterium number 〉=5.0 * 10
9The seed culture fluid of cfu/ml;
Yeast (
Pichia fermentansLodder) ACCC 20319
TThe preparation of seed culture fluid: with yeast (
Pichia fermentansLodder) ACCC 20319
TUnder aseptic condition, be inoculated in the liquid nutrient medium, described liquid nutrient medium is: the content of each component in 1.0L distilled water is: anhydrous magnesium sulfate 1.0g, potassium primary phosphate 2.0g, urea 3.0g, corn steep liquor 1.0g, manganous sulfate 0.02g, ferrous sulfate 0.2g, the pH value transfers to 5.5, cultivated 4～5 days, and obtained yeast bacterium number 〉=0.1 * 10 for 30 ℃-31 ℃
9The seed culture fluid of cfu/ml;
Bacillus natto (
Bacillus natto Swamura) seed culture fluid of ACCC10614 and subtilis (
Bacillus subtillsSubsp.
SpizieniiNakamura et al) ACCC 10242
TThe preparation of seed culture fluid: with Bacillus natto (
Bacillus natto Swamura) ACCC 10614 and subtilis (
Bacillus subtillsSubsp.
SpizieniiNakamura et al) ACCC 10242
TUnder aseptic condition, be inoculated in the liquid nutrient medium respectively, described liquid nutrient medium is: the content of each component in 1.0L distilled water is: peptone 10.0g, extractum carnis 3.0g, sodium-chlor 5.0g, the pH value transfers to 7.0,28 ℃～32 ℃, single spawn culture 7～8 days obtains Bacillus natto bacterium number 〉=5.0 * 10 respectively
9The Bacillus natto seed culture fluid of cfu/ml and bacterial count of bacillus subtilis 〉=5.0 * 10
9The subtilis seed culture fluid of cfu/ml;
Lactobacillus (
Lactococcus lactis) seed culture fluid of ACCC 11092 and bifidus bacillus (
BifidobacteriumSp) preparation of the seed culture fluid of ACCC 11054: with lactobacillus (
Lactococcuslactis) ACCC 11092 and bifidus bacillus (
BifidobacteriumSp) ACCC 11054 is inoculated in the liquid nutrient medium under aseptic condition respectively, described liquid nutrient medium is: the content of each component in 1.0L distilled water is: tomato juice 200.0ml, Zulkovsky starch 0.5g, peptone 15.0g, sodium-chlor 5.0g, yeast powder 6.0g, tween 80 1.0g, glucose 20.0g, the pH value transfers to 7.2,30 ℃～35 ℃ single spawn culture 2～3 days obtain lactobacillus bacterium number 〉=5.0 * 10 respectively
9The lactobacillus seed nutrient solution of cfu/ml and bifidus bacillus bacterium number 〉=5.0 * 10
9The bifidus bacillus seed nutrient solution of cfu/ml;
(2) preparation of many bacterial classifications of used for aquiculture microbial compound preparation:
Respectively the photosynthetic bacterium that step (1) is prepared (
Rhodoplanes elegans) ACCC 10649, yeast (
Pichia fermentansLodder) ACCC 20319
T, Bacillus natto (
Bacillus natto Swamura) ACCC10614, subtilis (
Bacillus subtillsSubsp.
SpizieniiNakamura et al) ACCC 10242
T, lactobacillus (
Lactococcus lactis) ACCC 11092 and bifidus bacillus (
BifidobacteriumSp) seed culture fluid of ACCC 11054 by its weight percentage is: the seed culture fluid 25%～35% of photosynthetic bacterium, saccharomycetic seed culture fluid 5%～15%, the seed culture fluid 15%～25% of Bacillus natto, the seed culture fluid 15%～25% of subtilis, the seed culture fluid 5%～15% of lactobacillus, the seed culture fluid 5%～15% of bifidus bacillus, under aseptic condition, be inoculated in the same liquid nutrient medium respectively, inoculum size is 1 ﹪～2 ﹪ of liquid nutrient medium weight, the content of each component in 1.0L distilled water is in the described liquid nutrient medium: brown sugar 50.0g, honey 10.0g, at oxygen dissolved is that 5 ﹪ are that the volume ratio of the volume of oxygen in the fermentor tank and fermented liquid is under 5% the condition, 28 ℃～32 ℃ closeds leave standstill cultivated 7～15 days, made the pH value drop to 3.0-3.5 and formed.
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| CN114456909B (en) * | 2022-03-17 | 2022-09-06 | 广东工业大学 | Deep-sea microorganism separation culture device and culture method |
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