CN101974434B - 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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- 238000002360 preparation method Methods 0.000 title claims abstract description 68
- 238000009360 aquaculture Methods 0.000 title claims abstract description 36
- 244000144974 aquaculture Species 0.000 title claims abstract description 29
- 230000000813 microbial effect Effects 0.000 title claims abstract description 27
- 150000001875 compounds Chemical class 0.000 title claims abstract description 23
- 238000011218 seed culture Methods 0.000 claims abstract description 114
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 36
- 235000014897 Streptococcus lactis Nutrition 0.000 claims abstract description 32
- 238000000855 fermentation Methods 0.000 claims abstract description 25
- 230000004151 fermentation Effects 0.000 claims abstract description 25
- 230000001580 bacterial effect Effects 0.000 claims abstract description 16
- 241000186000 Bifidobacterium Species 0.000 claims abstract description 3
- 239000012531 culture fluid Substances 0.000 claims description 110
- 241000894006 Bacteria Species 0.000 claims description 61
- 239000007788 liquid Substances 0.000 claims description 44
- 235000015097 nutrients Nutrition 0.000 claims description 44
- 244000025254 Cannabis sativa Species 0.000 claims description 34
- 210000000481 breast Anatomy 0.000 claims description 33
- 241000235648 Pichia Species 0.000 claims description 31
- 244000057717 Streptococcus lactis Species 0.000 claims description 31
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 31
- 230000000243 photosynthetic effect Effects 0.000 claims description 30
- 239000012153 distilled water Substances 0.000 claims description 17
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 11
- 239000001301 oxygen Substances 0.000 claims description 11
- 229910052760 oxygen Inorganic materials 0.000 claims description 11
- 239000001888 Peptone Substances 0.000 claims description 9
- 108010080698 Peptones Proteins 0.000 claims description 9
- 240000004808 Saccharomyces cerevisiae Species 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
- 241000194041 Lactococcus lactis subsp. lactis Species 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
- 235000014969 Streptococcus diacetilactis Nutrition 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
- 241000521553 Pichia fermentans Species 0.000 abstract description 3
- 201000010099 disease Diseases 0.000 abstract description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 3
- 244000063299 Bacillus subtilis Species 0.000 abstract description 2
- 235000014469 Bacillus subtilis Nutrition 0.000 abstract description 2
- 241000194035 Lactococcus lactis Species 0.000 abstract 1
- 241001148165 Rhodoplanes elegans Species 0.000 abstract 1
- 238000012258 culturing Methods 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
- 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
- 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
- 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
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-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
- 210000004102 animal cell Anatomy 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit 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
- 230000008859 change 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
- 238000012851 eutrophication Methods 0.000 description 1
- 239000004459 forage Substances 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000000968 intestinal effect Effects 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
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 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
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- 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
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- 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 multi-bacterium microbial compound preparations for aquaculture and preparation method thereof.
Background technology
A large amount of appearance along with the mass-producing culture fishery; cultivation density constantly increases; movement in the aquaculture water and the residue of bait constantly increase; the pathogenic microorganism amount reproduction; the aggravation of aquaculture water Eutrophic Extent; ecotope suffers serious destruction; this causes aquatic animal on a large scale and all kinds of diseases and emergency rating frequently occur; 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 used in the existing institute of aquaculture, still also exists many problems.The most of microbe preparation is only brought into play effect for aquaculture water, but aquatic animal itself is not had direct effect.
Summary of the invention
Technical problem to be solved by this invention provides a kind of multi-bacterium microbial compound preparations for aquaculture and preparation method thereof.This multi-bacterium microbial compound preparations for aquaculture 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 multi-bacterium microbial compound preparations for aquaculture is characterized in that: it be by photosynthetic bacterium (
Rhodopaeudomonas palustris ) ACCC 10649, the fermentation pichia spp (
Pichia fermentans Lodder) ACCC 20319
T, subtilis withered grass subspecies (
Bacillus subtills Subsp.
Subtilis Cohn) ACCC 10614, subtilis Si Shi subspecies (
Bacillus subtills Subsp.
Spizienii ) ACCC 10242
T, Lactococcus lactis breast subspecies (
Lactococcus lactis Subsp.
Lactis ) ACCC 11092 and bifidus bacillus (
Bifidobacterium Sp) ACCC 11054 carries out respectively the seed culture fluid that single spawn culture obtains and is inoculated in the same substratum, then carry out fermentor cultivation, the pH value for the treatment of nutrient solution naturally is down to 3.0～3.5 and is obtained the bacterium number of photosynthetic bacterium ACCC 10649 〉=5.0 * 10 in the said preparation
9Cfu/ml, fermentation pichia spp ACCC 20319
TBacterium number 〉=0.1 * 10
9Cfu/ml, the bacterium number of subtilis withered grass subspecies ACCC 10614 〉=5.0 * 10
9Cfu/ml, subtilis Si Shi subspecies ACCC 10242
TBacterium number 〉=5.0 * 10
9Cfu/ml, the bacterium number of Lactococcus lactis breast subspecies ACCC 11092 〉=5.0 * 10
9Cfu/ml, the bacterium number of bifidus bacillus ACCC 11054 〉=5.0 * 10
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, the seed culture fluid 5%～15% of fermentation pichia spp, the seed culture fluid 15%～25% of subtilis withered grass subspecies, the seed culture fluid 15%～25% of subtilis Si Shi subspecies, the seed culture fluid 5%～15% of Lactococcus lactis breast subspecies, the seed culture fluid 5%～15% of bifidus bacillus.
A kind of preparation method of multi-bacterium microbial compound preparations for aquaculture is characterized in that: comprise the steps:
(1) preparation of seed culture fluid:
The preparation of the seed culture fluid of photosynthetic bacterium ACCC 10649: photosynthetic bacterium 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;
Fermentation pichia spp ACCC 20319
TThe preparation of seed culture fluid: pichia spp ACCC 20319 will ferment
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 for 30 ℃-31 ℃, Pichia yeast number 〉=0.1 * 10 obtains fermenting
9The seed culture fluid of cfu/ml;
The seed culture fluid of subtilis withered grass subspecies ACCC 10614 and subtilis Si Shi subspecies ACCC 10242
TThe preparation of seed culture fluid: with subtilis withered grass subspecies ACCC 10614 and subtilis Si Shi subspecies 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 respectively subtilis withered grass subspecies bacterium number 〉=5.0 * 10
9The subtilis withered grass subspecies seed culture fluid of cfu/ml and subtilis Si Shi subspecies bacterium number 〉=5.0 * 10
9The subtilis Si Shi subspecies seed culture fluid of cfu/ml;
The preparation of the seed culture fluid of Lactococcus lactis breast subspecies ACCC 11092 and the seed culture fluid of bifidus bacillus ACCC 11054: Lactococcus lactis breast subspecies ACCC 11092 and bifidus bacillus ACCC 11054 are 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 respectively Lactococcus lactis breast subspecies bacterium number 〉=5.0 * 10
9Lactococcus lactis breast subspecies seed culture fluid and bifidus bacillus bacterium number 〉=5.0 * 10 of cfu/ml
9The bifidus bacillus seed nutrient solution of cfu/ml;
(2) preparation of multi-bacterium microbial compound preparations for aquaculture:
The photosynthetic bacterium ACCC 10649 that respectively step (1) is prepared, fermentation pichia spp ACCC 20319
T, subtilis withered grass subspecies ACCC10614, subtilis Si Shi subspecies ACCC 10242
TThe seed culture fluid of Lactococcus lactis breast subspecies ACCC 11092 and bifidus bacillus ACCC 11054 by its weight percentage is: the seed culture fluid 25%～35% of photosynthetic bacterium, the seed culture fluid 5%～15% of fermentation pichia spp, the seed culture fluid 15%～25% of subtilis withered grass subspecies, the seed culture fluid 15%～25% of subtilis Si Shi subspecies, the seed culture fluid 5%～15% of Lactococcus lactis breast subspecies, the seed culture fluid 5%～15% of bifidus bacillus, under aseptic condition, be inoculated into respectively in the same liquid nutrient medium, 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, be that 5 ﹪ are that the volume ratio of the volume of oxygen in the fermentor tank and fermented liquid is under 5% the condition at oxygen dissolved, 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 there is the phenomenon that mutually rejects, seizes Tan Yuan ∕ nitrogenous source and mutually corrode in different strain microorganism compound tense between each bacterial classification.The present invention is fermented the bacterial classification of different qualities respectively, obtain seed culture fluid, then be inoculated in respectively by a certain percentage mixed culture in the same substratum, 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 can avoid the mutual rejection that exists between the different strain, the phenomenon of seizing Tan ∕ nitrogenous source and mutually corroding.In use, the used for aquiculture microbial compound preparation of the present invention preparation put into the cultivation waters after because the change of environment can be so that 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 ACCC 10649: consume unnecessary nutritive substance in the water body, produce a large amount of useful nutritive substances to aquatic animal, reduce the objectionable impuritiess such as ammonia nitrogen, sulfide.
(2) fermentation pichia spp ACCC 20319
T: produce in a large number the essential multiple amino acids of animal.
(3) subtilis Si Shi subspecies ACCC 10242
T: produce proteolytic enzyme, amylase and subtilyne.
(4) Lactococcus lactis breast subspecies 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 ACCC 11054: safeguard enteron aisle normal bacteria colony balance, suppress the growth of pathogenic bacteria, synthesise vitamins, amino acid and raising body produce VITMAIN B1, B2, B6, B12 and L-Ala, α-amino-isovaleric acid, aspartic acid and Threonine etc. to the absorption of calcium ion in enteron aisle.
(6) subtilis withered grass subspecies ACCC 10614: the regulating intestinal canal flora, 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.Lactococcus lactis breast subspecies ACCC 11092 in this preparation, bifidus bacillus ACCC 11054, subtilis withered grass subspecies ACCC 10614 can grow rapidly in the enteron aisle of aquatic animal surely, set up the population advantage of probiotics, promote the dietetic alimentation of aquatic animal, produce multiple aquatic animal grow necessary amino acid, VITAMIN and organic acid, and bifidus bacillus ACCC 11054, subtilis withered grass subspecies ACCC10614 be the immunologic function of enhancing body significantly, effectively prevents digestive tract infection, reduces emergency rating;
2. purifying aquatic water.Photosynthetic bacterium ACCC 10649 in this preparation, fermentation pichia spp ACCC 20319
TWith subtilis Si Shi subspecies 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 rich nutrition level of aquaculture water, improve the oxygen dissolved of water body, produce simultaneously multiple proteins, the natural bait of high-quality is provided for aquatic animal, 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 for the 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:
The preparation of the seed culture fluid of photosynthetic bacterium (Rhodopaeudomonas palustris) ACCC 10649: with photosynthetic bacterium (Rhodopaeudomonas palustris) ACCC 10649 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.
Fermentation pichia spp ACCC 20319
TThe preparation of seed culture fluid: pichia spp ACCC 20319 will ferment
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 for 30 ℃-31 ℃, Pichia yeast number 〉=0.1 * 10 obtains fermenting
9The seed culture fluid of cfu/ml.
Subtilis withered grass subspecies ACCC 10614 seed culture fluids and subtilis Si Shi subspecies ACCC 10242
TThe preparation of seed culture fluid: with subtilis withered grass subspecies ACCC 10614 and subtilis Si Shi subspecies 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 respectively subtilis withered grass subspecies bacterium number 〉=5.0 * 10
9The subtilis withered grass subspecies seed culture fluid of cfu/ml and subtilis Si Shi subspecies bacterium number 〉=5.0 * 10
9The subtilis Si Shi subspecies seed culture fluid of cfu/ml.
The preparation of Lactococcus lactis breast subspecies ACCC 11092 seed culture fluids and bifidus bacillus ACCC 11054 seed culture fluids: under aseptic condition, be inoculated in the same liquid nutrient medium Lactococcus lactis breast subspecies ACCC 11092 and bifidus bacillus ACCC 11054 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 respectively Lactococcus lactis breast subspecies bacterium number 〉=5.0 * 10
9Lactococcus lactis breast subspecies seed culture fluid and bifidus bacillus bacterium number 〉=5.0 * 10 of cfu/ml
9The bifidus bacillus seed nutrient solution of cfu/ml.
(2) preparation of used for aquiculture microbial preparation:
The seed culture fluid of each bacterial classification that respectively step (1) is prepared according to the seed culture fluid of the seed culture fluid of photosynthetic bacterium ACCC 10649, fermentation pichia spp ACCC 20319T, subtilis withered grass subspecies (
Bacillus natto Swamura) seed culture fluid of ACCC10614, the seed culture fluid of subtilis Si Shi subspecies ACCC 10242T, Lactococcus lactis breast subspecies (
Lactococcus lactis) weight percentage of the seed culture fluid of ACCC 11092 and the seed culture fluid of bifidus bacillus ACCC 11054 is: the seed culture fluid 25% of photosynthetic bacterium, the seed culture fluid 5% of fermentation pichia spp, the seed culture fluid 15% of subtilis withered grass subspecies, the seed culture fluid 25% of subtilis Si Shi subspecies, the seed culture fluid 15% of Lactococcus lactis breast subspecies, the seed culture fluid 15% of bifidus bacillus, under aseptic condition, be inoculated into respectively in the same liquid nutrient medium, 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 naturally drop to 3.0-3.5 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: the bacterium number of photosynthetic bacterium ACCC 10649 〉=5.0 * 10
9Cfu/ml, fermentation pichia spp ACCC 20319
TBacterium number 〉=〉=0.1 * 10
9Cfu/ml, the bacterium number of subtilis withered grass subspecies ACCC 10614 〉=5.0 * 10
9Cfu/ml, subtilis Si Shi subspecies ACCC 10242
TBacterium number 〉=5.0 * 10
9Cfu/ml, the bacterium number of Lactococcus lactis breast subspecies ACCC 11092 〉=5.0 * 10
9Cfu/ml, the bacterium number of bifidus bacillus ACCC 11054 〉=5.0 * 10
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:
The seed culture fluid of each bacterial classification that respectively step (1) is prepared is according to seed culture fluid, the fermentation pichia spp ACCC 20319 of photosynthetic bacterium ACCC 10649
TSeed culture fluid, the seed culture fluid of subtilis withered grass subspecies ACCC10614, subtilis Si Shi subspecies ACCC 10242
TSeed culture fluid, Lactococcus lactis breast subspecies (
Lactococcus lactis Subsp.
Lactis ) weight percentage of the seed culture fluid of ACCC 11092 and the seed culture fluid of bifidus bacillus ACCC 11054 is: the seed culture fluid 35% of photosynthetic bacterium, the seed culture fluid 15% of fermentation pichia spp, the seed culture fluid 25% of subtilis withered grass subspecies, the seed culture fluid 15% of subtilis Si Shi subspecies, the seed culture fluid 5% of Lactococcus lactis breast subspecies, the seed culture fluid 5% of bifidus bacillus, under aseptic condition, be inoculated into respectively in the same liquid nutrient medium, 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 naturally drop to 3.0～3.5 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: the bacterium number of photosynthetic bacterium ACCC 10649 〉=5.0 * 10
9Cfu/ml, fermentation pichia spp ACCC 20319
TBacterium number 〉=0.1 * 10
9Cfu/ml, the bacterium number of subtilis withered grass subspecies ACCC 10614 〉=5.0 * 10
9Cfu/ml, subtilis Si Shi subspecies ACCC 10242
TBacterium number 〉=5.0 * 10
9Cfu/ml, the bacterium number of Lactococcus lactis breast subspecies ACCC 11092 〉=5.0 * 10
9Cfu/ml, the bacterium number of bifidus bacillus ACCC 11054 〉=5.0 * 10
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:
The seed culture fluid of each bacterial classification that respectively step (1) is prepared is according to seed culture fluid, the fermentation pichia spp ACCC 20319 of photosynthetic bacterium ACCC 10649
TSeed culture fluid, the seed culture fluid of subtilis withered grass subspecies ACCC 10614, subtilis Si Shi subspecies ACCC 10242
TSeed culture fluid, the weight percentage of the seed culture fluid of Lactococcus lactis breast subspecies ACCC 11092 and the seed culture fluid of bifidus bacillus ACCC 11054 is: the seed culture fluid 30% of photosynthetic bacterium, the seed culture fluid 10% of fermentation pichia spp, the seed culture fluid 20% of subtilis withered grass subspecies, the seed culture fluid 20% of subtilis Si Shi subspecies, the seed culture fluid 10% of Lactococcus lactis breast subspecies, the seed culture fluid 10% of bifidus bacillus, under aseptic condition, be inoculated into respectively in the same liquid nutrient medium, 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 naturally drop to 3.0～3.5 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: the bacterium number of photosynthetic bacterium ACCC 10649 〉=5.0 * 10
9Cfu/ml, fermentation pichia spp ACCC 20319
TBacterium number 〉=0.1 * 10
9Cfu/ml, the bacterium number of subtilis withered grass subspecies ACCC 10614 〉=5.0 * 10
9Cfu/ml, subtilis Si Shi subspecies ACCC 10242
TBacterium number 〉=5.0 * 10
9Cfu/ml, the bacterium number of Lactococcus lactis breast subspecies ACCC 11092 〉=5.0 * 10
9Cfu/ml, the bacterium number of bifidus bacillus ACCC 11054 〉=5.0 * 10
9Cfu/ml.
Application Example:
Adopt the aquaculture microbial compound preparation of the embodiment of the invention 1 preparation, in shrimp pond and fishpond, be implemented as follows respectively:
Take the shrimp pond as 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 front 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 contrast pond in test shrimp pond increases by 85%; Ammonia-nitrogen content≤0.1mg/mL, contrast shrimp pond on average descends 65%; Gross output contrast shrimp pond increases 25kg, amplification 15%; Commodity rate reaches 90%, and specification is large, and price is high, and contrast pond commodity rate is 54.5%, and specification is little, and price is low relatively.
Take in the fishpond as 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 front 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 contrast fishpond in test fishpond increases 100kg, amplification 25%; 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 (2)
1. multi-bacterium microbial compound preparations for aquaculture is characterized in that: it be by photosynthetic bacterium (
Rhodopaeudomonas palustris ) ACCC 10649, the fermentation pichia spp (
Pichia fermentanslodder ) ACCC 20319
T, subtilis withered grass subspecies (
Bacillus subtills Subsp.
Subtilis Cohn) ACCC 10614, subtilis Si Shi subspecies (
Bacillus subtills Subsp.
Spizienii ) ACCC 10242
T, Lactococcus lactis breast subspecies (
Lactococcus lactis Subsp.
Lactis ACCC 11092 and bifidus bacillus (
Bifidobacterium Sp) ACCC 11054 carries out respectively the seed culture fluid that single spawn culture obtains and is inoculated in the same substratum, then carry out fermentor cultivation, the pH value for the treatment of nutrient solution naturally is down to 3.0～3.5 and is obtained the bacterium number of photosynthetic bacterium ACCC 10649 〉=5.0 * 10 in the said preparation
9Cfu/ml, fermentation pichia spp ACCC 20319
TBacterium number 〉=0.1 * 10
9Cfu/ml, the bacterium number of subtilis withered grass subspecies ACCC 10614 〉=5.0 * 10
9Cfu/ml, subtilis Si Shi subspecies ACCC 10242
TBacterium number 〉=5.0 * 10
9Cfu/ml, the bacterium number of Lactococcus lactis breast subspecies ACCC 11092 〉=5.0 * 10
9Cfu/ml, the bacterium number of bifidus bacillus ACCC 11054 〉=5.0 * 10
9Cfu/ml;
The weight percentage of the seed culture fluid of described each bacterial classification is: the seed culture fluid 25%～35% of photosynthetic bacterium, the seed culture fluid 5%～15% of fermentation pichia spp, the seed culture fluid 15%～25% of subtilis withered grass subspecies, the seed culture fluid 15%～25% of subtilis Si Shi subspecies, the seed culture fluid 5%～15% of Lactococcus lactis breast subspecies, the seed culture fluid 5%～15% of bifidus bacillus.
2. the preparation method of multi-bacterium microbial compound preparations for aquaculture according to claim 1 is characterized in that: comprise the steps:
(1) preparation of seed culture fluid:
The preparation of the seed culture fluid of photosynthetic bacterium ACCC 10649: photosynthetic bacterium 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;
Fermentation pichia spp ACCC 20319
TThe preparation of seed culture fluid: pichia spp ACCC 20319 will ferment
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 for 30 ℃-31 ℃, Pichia yeast number 〉=0.1 * 10 obtains fermenting
9The seed culture fluid of cfu/ml;
The seed culture fluid of subtilis withered grass subspecies ACCC 10614 and subtilis Si Shi subspecies ACCC 10242
TThe preparation of seed culture fluid: with subtilis withered grass subspecies ACCC 10614 and subtilis Si Shi subspecies 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 respectively subtilis withered grass subspecies bacterium number 〉=5.0 * 10
9The subtilis withered grass subspecies seed culture fluid of cfu/ml and subtilis Si Shi subspecies bacterium number 〉=5.0 * 10
9The subtilis Si Shi subspecies seed culture fluid of cfu/ml;
The seed culture fluid of Lactococcus lactis breast subspecies ACCC 11092 and bifidus bacillus (
BifidobacteriumSp) preparation of the seed culture fluid of ACCC 11054: with Lactococcus lactis breast subspecies 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 respectively Lactococcus lactis breast subspecies bacterium number 〉=5.0 * 10
9Lactococcus lactis breast subspecies seed culture fluid and bifidus bacillus bacterium number 〉=5.0 * 10 of cfu/ml
9The bifidus bacillus seed nutrient solution of cfu/ml;
(2) preparation of multi-bacterium microbial compound preparations for aquaculture:
The photosynthetic bacterium ACCC 10649 that respectively step (1) is prepared, fermentation pichia spp ACCC 20319
T, subtilis withered grass subspecies ACCC10614, subtilis Si Shi subspecies ACCC 10242
TThe seed culture fluid of Lactococcus lactis breast subspecies ACCC 11092 and bifidus bacillus ACCC 11054 by its weight percentage is: the seed culture fluid 25%～35% of photosynthetic bacterium, the seed culture fluid 5%～15% of fermentation pichia spp, the seed culture fluid 15%～25% of subtilis withered grass subspecies, the seed culture fluid 15%～25% of subtilis Si Shi subspecies, the seed culture fluid 5%～15% of Lactococcus lactis breast subspecies, the seed culture fluid 5%～15% of bifidus bacillus, under aseptic condition, be inoculated into respectively in the same liquid nutrient medium, 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, the volume ratio of the volume of oxygen and fermented liquid is under 5% the condition in fermentor tank, 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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| CN103652329A (en) * | 2013-12-18 | 2014-03-26 | 大连金砣水产食品有限公司 | Compound probiotic biological agent |
| CN104770596A (en) * | 2015-04-07 | 2015-07-15 | 中国水产科学研究院珠江水产研究所 | Aquatic animal intestinal regulator and application thereof |
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| CN107574136B (en) * | 2017-10-30 | 2020-08-21 | 中国水产科学研究院淡水渔业研究中心 | Preparation method of semisolid microecological preparation for aquaculture |
| CN109810914A (en) * | 2018-04-19 | 2019-05-28 | 湛江恒兴养殖技术服务有限公司 | A kind of fishes and shrimps microbial compound preparation and preparation method thereof |
| CN108813223A (en) * | 2018-07-03 | 2018-11-16 | 珠海市西科水产养殖有限公司 | A kind of aquaculture specific complex microorganism formulation and preparation method thereof |
| CN109652331A (en) * | 2019-01-03 | 2019-04-19 | 沧州临港海益水产养殖有限公司 | For regulating and controlling the probiotics of pH value and the method for administration in sea-farming |
| CN110881442A (en) * | 2019-12-11 | 2020-03-17 | 湖南蒙生生物科技股份有限公司 | Method for producing ecological soft-shelled turtles by using herbal biological conditioning powder |
| CN111793575A (en) * | 2020-06-03 | 2020-10-20 | 惠州学院 | Complex microbial inoculant and application thereof in aquaculture |
| CN113213642B (en) * | 2021-04-01 | 2023-03-17 | 浙江理工大学 | Water-regulating microecological preparation for micropterus salmoides seedling culture and preparation method thereof |
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