CN114045228B - Microbial water quality regulator and preparation method and application thereof - Google Patents
Microbial water quality regulator and preparation method and application thereof Download PDFInfo
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- CN114045228B CN114045228B CN202111136718.XA CN202111136718A CN114045228B CN 114045228 B CN114045228 B CN 114045228B CN 202111136718 A CN202111136718 A CN 202111136718A CN 114045228 B CN114045228 B CN 114045228B
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 117
- 230000000813 microbial effect Effects 0.000 title claims abstract description 32
- 238000002360 preparation method Methods 0.000 title abstract description 17
- 239000006041 probiotic Substances 0.000 claims abstract description 100
- 235000018291 probiotics Nutrition 0.000 claims abstract description 100
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims abstract description 80
- 241000894006 Bacteria Species 0.000 claims abstract description 77
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 47
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 47
- 241000190950 Rhodopseudomonas palustris Species 0.000 claims abstract description 45
- 241000235342 Saccharomycetes Species 0.000 claims abstract description 43
- 239000004310 lactic acid Substances 0.000 claims abstract description 40
- 235000014655 lactic acid Nutrition 0.000 claims abstract description 40
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 37
- 241000195649 Chlorella <Chlorellales> Species 0.000 claims abstract description 29
- 230000001546 nitrifying effect Effects 0.000 claims abstract description 29
- 230000012010 growth Effects 0.000 claims abstract description 26
- 230000000529 probiotic effect Effects 0.000 claims abstract description 21
- 239000003337 fertilizer Substances 0.000 claims abstract description 20
- 238000000855 fermentation Methods 0.000 claims description 60
- 230000004151 fermentation Effects 0.000 claims description 60
- 239000000843 powder Substances 0.000 claims description 22
- 241000186660 Lactobacillus Species 0.000 claims description 19
- 238000004108 freeze drying Methods 0.000 claims description 19
- 229940039696 lactobacillus Drugs 0.000 claims description 19
- 239000003223 protective agent Substances 0.000 claims description 19
- 238000000034 method Methods 0.000 claims description 14
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 12
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 7
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 7
- 241001453382 Nitrosomonadales Species 0.000 claims description 6
- 230000001590 oxidative effect Effects 0.000 claims description 6
- 241000235646 Cyberlindnera jadinii Species 0.000 claims description 4
- 241000194031 Enterococcus faecium Species 0.000 claims description 4
- 244000199866 Lactobacillus casei Species 0.000 claims description 4
- 235000013958 Lactobacillus casei Nutrition 0.000 claims description 4
- 241000186840 Lactobacillus fermentum Species 0.000 claims description 4
- 240000006024 Lactobacillus plantarum Species 0.000 claims description 4
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims description 4
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims description 4
- IOVCWXUNBOPUCH-UHFFFAOYSA-N Nitrous acid Chemical compound ON=O IOVCWXUNBOPUCH-UHFFFAOYSA-N 0.000 claims description 4
- 229940017800 lactobacillus casei Drugs 0.000 claims description 4
- 229940012969 lactobacillus fermentum Drugs 0.000 claims description 4
- 229940072205 lactobacillus plantarum Drugs 0.000 claims description 4
- 239000008101 lactose Substances 0.000 claims description 4
- 235000020183 skimmed milk Nutrition 0.000 claims description 4
- 230000004913 activation Effects 0.000 claims description 3
- 150000003839 salts Chemical class 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 2
- 230000031877 prophase Effects 0.000 claims 2
- 238000009360 aquaculture Methods 0.000 abstract description 25
- 244000144974 aquaculture Species 0.000 abstract description 25
- 241001465754 Metazoa Species 0.000 abstract description 10
- 230000003698 anagen phase Effects 0.000 abstract description 3
- 239000001963 growth medium Substances 0.000 description 40
- 241000238557 Decapoda Species 0.000 description 18
- 238000012258 culturing Methods 0.000 description 18
- 238000009630 liquid culture Methods 0.000 description 14
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 12
- 230000000052 comparative effect Effects 0.000 description 12
- 239000007787 solid Substances 0.000 description 12
- 239000012153 distilled water Substances 0.000 description 10
- 238000005303 weighing Methods 0.000 description 10
- 244000005700 microbiome Species 0.000 description 9
- 230000001580 bacterial effect Effects 0.000 description 8
- 238000005286 illumination Methods 0.000 description 8
- 230000008901 benefit Effects 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- 229920001817 Agar Polymers 0.000 description 6
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 6
- 239000001888 Peptone Substances 0.000 description 6
- 108010080698 Peptones Proteins 0.000 description 6
- 239000008272 agar Substances 0.000 description 6
- 239000001110 calcium chloride Substances 0.000 description 6
- 229910001628 calcium chloride Inorganic materials 0.000 description 6
- 229940041514 candida albicans extract Drugs 0.000 description 6
- 238000012136 culture method Methods 0.000 description 6
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 6
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 6
- 235000019341 magnesium sulphate Nutrition 0.000 description 6
- 238000010907 mechanical stirring Methods 0.000 description 6
- 235000019319 peptone Nutrition 0.000 description 6
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 6
- 239000011780 sodium chloride Substances 0.000 description 6
- 239000012138 yeast extract Substances 0.000 description 6
- 230000000694 effects Effects 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 230000004083 survival effect Effects 0.000 description 5
- 241000195493 Cryptophyta Species 0.000 description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 4
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 4
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 4
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 4
- 235000019796 monopotassium phosphate Nutrition 0.000 description 4
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 4
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 description 4
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 description 4
- 230000003068 static effect Effects 0.000 description 4
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 2
- 235000011130 ammonium sulphate Nutrition 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 2
- 239000004327 boric acid Substances 0.000 description 2
- 239000004202 carbamide Substances 0.000 description 2
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 2
- 229910000366 copper(II) sulfate Inorganic materials 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 2
- 235000019797 dipotassium phosphate Nutrition 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 2
- 229960002413 ferric citrate Drugs 0.000 description 2
- 239000011790 ferrous sulphate Substances 0.000 description 2
- 235000003891 ferrous sulphate Nutrition 0.000 description 2
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 2
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 2
- NPFOYSMITVOQOS-UHFFFAOYSA-K iron(III) citrate Chemical compound [Fe+3].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NPFOYSMITVOQOS-UHFFFAOYSA-K 0.000 description 2
- 229910001629 magnesium chloride Inorganic materials 0.000 description 2
- 229940099596 manganese sulfate Drugs 0.000 description 2
- 239000011702 manganese sulphate Substances 0.000 description 2
- 235000007079 manganese sulphate Nutrition 0.000 description 2
- CNFDGXZLMLFIJV-UHFFFAOYSA-L manganese(II) chloride tetrahydrate Chemical compound O.O.O.O.[Cl-].[Cl-].[Mn+2] CNFDGXZLMLFIJV-UHFFFAOYSA-L 0.000 description 2
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000013535 sea water Substances 0.000 description 2
- 239000001632 sodium acetate Substances 0.000 description 2
- 235000017281 sodium acetate Nutrition 0.000 description 2
- 229910000029 sodium carbonate Inorganic materials 0.000 description 2
- 239000004317 sodium nitrate Substances 0.000 description 2
- 235000010344 sodium nitrate Nutrition 0.000 description 2
- 235000010288 sodium nitrite Nutrition 0.000 description 2
- RWVGQQGBQSJDQV-UHFFFAOYSA-M sodium;3-[[4-[(e)-[4-(4-ethoxyanilino)phenyl]-[4-[ethyl-[(3-sulfonatophenyl)methyl]azaniumylidene]-2-methylcyclohexa-2,5-dien-1-ylidene]methyl]-n-ethyl-3-methylanilino]methyl]benzenesulfonate Chemical compound [Na+].C1=CC(OCC)=CC=C1NC1=CC=C(C(=C2C(=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C)C=2C(=CC(=CC=2)N(CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C)C=C1 RWVGQQGBQSJDQV-UHFFFAOYSA-M 0.000 description 2
- 230000009044 synergistic interaction Effects 0.000 description 2
- 241000194108 Bacillus licheniformis Species 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- 241000192700 Cyanobacteria Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 238000012364 cultivation method Methods 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 238000013139 quantization Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000035882 stress Effects 0.000 description 1
- 230000004584 weight gain Effects 0.000 description 1
- 235000019786 weight gain Nutrition 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/32—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae
- C02F3/322—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae use of algae
- C02F3/325—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae use of algae as symbiotic combination of algae and bacteria
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/341—Consortia of bacteria
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/347—Use of yeasts or fungi
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/04—Preserving or maintaining viable microorganisms
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/12—Unicellular algae; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/18—Baker's yeast; Brewer's yeast
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/20—Nature of the water, waste water, sewage or sludge to be treated from animal husbandry
Abstract
The invention relates to the technical field of aquaculture, in particular to a microbial water quality regulator and a preparation method and application thereof. The microbial water quality regulator comprises a fertilizer-water phase probiotic agent, a growth phase probiotic agent and a culture later phase probiotic agent; the probiotics for the fertilizer and water period comprises 10-40% of lactic acid bacteria, 10-50% of chlorella and 10-50% of saccharomycetes by mass percent; the probiotics in the growth period comprises 10-40% of lactic acid bacteria, 10-40% of rhodopseudomonas palustris, 10-40% of saccharomycetes and 10-40% of bacillus subtilis by mass percent; the probiotics agent for the later period of cultivation comprises, by mass, 10-40% of lactic acid bacteria, 10-40% of rhodopseudomonas palustris, 10-40% of bacillus subtilis and 10-40% of nitrifying bacteria. The invention is respectively applicable to the early stage, the middle stage and the later stage of aquaculture by adjusting different types of probiotics. And (3) corresponding probiotics are added in different aquaculture periods, so that the water body requirements of different aquaculture periods are respectively ensured, a high-quality living environment is provided for aquatic animals, and the product quality is improved.
Description
Technical Field
The invention relates to the technical field of aquaculture, in particular to a microbial water quality regulator and a preparation method and application thereof.
Background
The whole aquaculture production process is not separated from water, and the advantages and disadvantages of the survival medium, namely water, directly determine whether aquaculture can benefit. Good water bodies comprise four key words of fertilizer, activity, tenderness and freshness. The fertilizer is enough in quantity of phytoplankton in water, and can provide natural feed for young cultured animals; the living is that the biological system in the water body is complete, three roles of a producer, a consumer and a decomposer normally play roles, and plants such as algae and the like in the water body can normally perform photosynthesis, and the ecological system is normal; tender refers to that phytoplankton and fungus in water are in an increased period, so that the cell state is good and the aging degree is low; "refreshing" means that the water quality is not turbid, and other impurities such as suspended matters are less. The good water body can promote the aquatic animals to fully absorb and convert the feed, improve the quality safety of the aquatic products and increase the aquaculture benefit. Therefore, the water quality regulation has important effect and significance.
The water quality regulator mainly comprises chemical preparations and microbial preparations. The chemical preparation mainly uses chemical or inorganic raw materials, the removal of nitrite and other pollutants in the water body depends on chemical or physical reactions, the use effect is not sustainable, an ecological organic circulation system is difficult to form, various chemicals are added in the water body, a temporary effect can be seen from indexes, but intense chemical and physical reactions can also generate unpredictable stress reactions on aquatic animals, scientific production of aquaculture is not facilitated, compared with the microbial water quality regulator, the ecological system can be better established, various water quality deterioration conditions can be effectively prevented and treated, and stable yield and high yield are realized. However, different microorganisms are mutually influenced and restricted, the microorganisms are extremely easy to be influenced by water temperature and pH, and the existing microorganism water quality regulator on the market has the problems of poor effect and the like.
Disclosure of Invention
In view of the above, it is necessary to provide a microbial water quality regulator, and a preparation method and application thereof. The microbial water quality regulator comprises three probiotics, and the probiotics are respectively applicable to the fertilizer water period, the growing period and the cultivation later period of aquaculture by adjusting different types of the probiotics. And (3) corresponding probiotics are added in different aquaculture periods, so that the water body requirements of different aquaculture periods are respectively ensured. Meanwhile, the probiotics of different types act on the same aquaculture period to realize synergistic interaction, so that the water quality of the aquaculture water body is regulated, the survival rate of aquatic animals is improved, and the aquaculture benefit is increased.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
in a first aspect, the present invention provides a microbial water quality regulator comprising a fertilizer-water phase probiotic, a growth phase probiotic, and a post-cultivation probiotic; the probiotics for the fertilizer and water period comprises 10-40% of lactic acid bacteria, 10-50% of chlorella and 10-50% of saccharomycetes by mass percent; the probiotics in the growth period comprises 10-40% of lactic acid bacteria, 10-40% of rhodopseudomonas palustris, 10-40% of saccharomycetes and 10-40% of bacillus subtilis by mass percent; the probiotics agent for the later period of cultivation comprises, by mass, 10-40% of lactic acid bacteria, 10-40% of rhodopseudomonas palustris, 10-40% of bacillus subtilis and 10-40% of nitrifying bacteria.
Preferably, the probiotics for the water-fertilizer period comprises 30% of lactic acid bacteria, 40% of chlorella and 30% of saccharomycetes in percentage by mass; the probiotics in the growth period comprises 20% of lactic acid bacteria, 20% of rhodopseudomonas palustris, 30% of saccharomycetes and 30% of bacillus subtilis in percentage by mass; the post-cultivation probiotics comprise 20% of lactic acid bacteria, 30% of rhodopseudomonas palustris, 30% of bacillus subtilis and 20% of nitrifying bacteria in percentage by mass.
Further, the lactic acid bacteria include lactobacillus fermentum, lactobacillus plantarum, lactobacillus casei and streptococcus faecium.
Further, the yeasts include Saccharomyces cerevisiae and Candida utilis.
Further, the nitrifying bacteria include ammonia oxidizing bacteria and nitrous acid oxidizing bacteria.
Further, the lactobacillus consists of 50-70% of lactobacillus fermentation liquor, 20-40% of shell powder and 10-30% of freeze-drying protective agent by mass percent; the rhodopseudomonas palustris consists of 50-70% of rhodopseudomonas palustris fermentation liquor, 20-40% of shell powder and 10-30% of freeze-drying protective agent by mass percent; the chlorella consists of 50-70% of chlorella fermentation liquor, 20-40% of shell powder and 10-30% of freeze-drying protective agent by mass percent; the saccharomycete consists of 50-70% saccharomycete fermentation liquor, 20-40% shell powder and 10-30% freeze-drying protective agent in percentage by mass; the bacillus subtilis consists of 50-70% of bacillus subtilis fermentation liquor, 20-40% of shell powder and 10-30% of freeze-drying protective agent by mass percent; the nitrifying bacteria consists of 50-70% of nitrifying bacteria fermentation liquor, 20-40% of shell powder and 10-30% of freeze-drying protective agent by mass percent.
Further, the freeze-drying protective agent consists of 50-80% of skimmed milk powder, 2-20% of lactose and 10-30% of glycerol in percentage by mass.
Preferably, the lyoprotectant consists of 75% skimmed milk powder, 5% lactose and 20% glycerol by mass percent.
Further, the preparation method of the lactobacillus fermentation broth comprises the following steps:
a1: culturing lactobacillus fermentum, lactobacillus plantarum, lactobacillus casei and streptococcus faecium in a slant solid culture medium at 28-30 ℃ for 36 hours to obtain a first-stage pure culture, then inoculating the first-stage pure culture into a 500mL triangular flask filled with a liquid culture medium for 24 hours, performing liquid shaking culture to obtain a second-stage pure culture, and finally inoculating the second-stage pure culture into a 1.8L mechanical stirring fermentation tank for culturing for 24 hours to obtain lactobacillus fermentation broth, wherein the bacterial count of the lactobacillus fermentation broth is more than or equal to 7log CFU/g; wherein the liquid culture medium is wort culture medium, and the solid culture medium is wort culture medium plus agar;
further, the culture method of the rhodopseudomonas palustris fermentation broth comprises the following steps:
b1: preparing a culture medium: 3.5g of sodium acetate, 1.0g of ammonia chloride, 1.2g of monopotassium phosphate, 0.4g of dipotassium phosphate, 0.1g of magnesium chloride, 0.1g of calcium chloride, 0.4g of yeast extract and 0.8g of peptone are weighed, and distilled water is used for constant volume to 1 liter; ph=7.0, autoclaving at 121 ℃ for 20 min;
b2: culturing rhodopseudomonas palustris in a test tube with a prepared culture medium at 28-30 ℃ for 96-144 hours under illumination to obtain a first-stage pure culture, then placing the first-stage pure culture in a 500mL triangular flask for 60-72 hours under constant temperature shaking culture to obtain a second-stage pure culture, and finally inoculating the second-stage pure culture into a 1.8L plastic flask for static culture under illumination at 28-30 ℃ to obtain rhodopseudomonas palustris fermentation broth, wherein the bacterial count of the rhodopseudomonas palustris fermentation broth is more than or equal to 7log CFU/g.
Further, the culture method of the chlorella fermentation broth comprises the following steps:
c1: preparing a culture medium: weighing 0.16g of sodium nitrate, 0.02g of monopotassium phosphate, 0.05g of magnesium sulfate, 0.04g of urea, 0.02g of calcium chloride, 0.02g of sodium bicarbonate, 1.00g of sodium chloride, 4.00mg of ferric citrate, 0.08mg of cupric sulfate pentahydrate, 2.86mg of boric acid, 1.81mg of manganese chloride tetrahydrate and 0.21mg of sodium molybdate dihydrate; the sea water is used for constant volume to 1 liter; ph=7.0, autoclaving at 121 ℃ for 20 min;
c2: culturing chlorella in a test tube filled with a culture medium at 23-24 ℃ for 96-144 hours under illumination to obtain a first-stage pure culture, then culturing in a 500mL triangular flask at constant temperature for 60-72 hours under shaking to obtain a second-stage pure culture, then inoculating to a 1.8L plastic bottle, and culturing at 23-24 ℃ under illumination for static condition to obtain chlorella fermentation broth, wherein the number of algae is more than or equal to 7log CFU/g.
Further, the method for culturing the saccharomycete fermentation broth comprises the following steps:
d1: preparing a liquid culture medium: weighing 5g of yeast extract, 10g of peptone and 20g of glucose, fixing the volume by distilled water by 1L, adjusting the pH to be 6.4, and autoclaving at 118 ℃ for 20 minutes; preparing a solid culture medium: adding 10g of agar on the basis of the liquid culture medium;
d2: and (3) carrying out slant solid culture on the saccharomyces cerevisiae and the candida utilis at 28-30 ℃ to obtain a first-stage pure culture, then inoculating the first-stage pure culture into a 500mL triangular flask filled with a liquid culture medium, carrying out constant-temperature shaking culture to obtain a second-stage pure culture, and finally inoculating the second-stage pure culture into a 1.8L mechanical stirring fermentation tank to culture for 24 hours to obtain a saccharomycete fermentation broth, wherein the number of bacteria of the saccharomycete fermentation broth is more than or equal to 8log CFU/g.
Further, the method for culturing the bacillus subtilis fermentation broth comprises the following steps:
e1: preparing a culture medium: weighing 10.0g of peptone, 5.0g of yeast extract, 10.0g of sodium chloride and 10.0g of glucose, fixing the volume to 1 liter with distilled water, adjusting the pH to be 7.0, and carrying out damp-heat autoclaving at 118 ℃ for 18 minutes; preparing a solid culture medium: adding 10g of agar on the basis of the liquid culture medium;
e2: performing inclined solid culture on bacillus subtilis at 28-30 ℃ to obtain a first-stage pure culture, then inoculating the first-stage pure culture into a 500mL triangular flask filled with a liquid culture medium, performing constant-temperature shaking culture to obtain a second-stage pure culture, and finally inoculating the second-stage pure culture into a 1.8L mechanical stirring fermentation tank to perform culture for 24 hours to obtain bacillus subtilis fermentation broth, wherein the bacterial count of the bacillus subtilis fermentation broth is more than or equal to 8log CFU/g.
Further, the culture method of the nitrifying bacteria fermentation liquor comprises the following steps:
f1: preparing an ammonia oxidizing bacteria culture medium: weighing 0.5g of ammonium sulfate, 0.3g of sodium chloride, 0.03g of ferrous sulfate, 1.0g of dipotassium hydrogen phosphate, 0.05g of magnesium sulfate, 7.5g of calcium chloride and distilled water to a constant volume of 1 liter; ph=7.0, autoclaving at 121 ℃ for 20 min;
f2: preparing a nitrous acid oxidizing bacteria culture medium: 1.0g of sodium nitrite, 0.03g of magnesium sulfate, 0.01g of manganese sulfate, 0.75g of dipotassium hydrogen phosphate, 1.0g of sodium carbonate, 0.25g of disodium hydrogen phosphate and distilled water to a constant volume of 1 liter are weighed; ph=7.0, autoclaving at 121 ℃ for 20 min;
f3: respectively culturing at 28deg.C under 160 r/min for 144 hr to obtain ammonia oxidizing bacteria fermentation broth and nitrite oxidizing bacteria fermentation broth, and mixing to obtain nitrifying bacteria fermentation broth with total bacterial count not less than 8log CFU/g.
In a second aspect, the invention provides a preparation method of a microbial water quality regulator, which comprises the steps of mixing lactobacillus, chlorella and saccharomycetes according to a proportion to obtain a probiotic agent in a fertilizer water period; mixing lactobacillus, rhodopseudomonas palustris, saccharomycetes and bacillus subtilis according to a proportion to obtain a probiotic agent in a growth period; and mixing lactobacillus, bacillus subtilis and nitrifying bacteria in proportion to obtain the probiotic agent in the later period of cultivation.
In a third aspect, the invention provides an application method of a microbial water quality regulator, wherein in the aquaculture process, a fertilizer-water phase probiotic is used in the early stage of cultivation, a growth phase probiotic is used in the middle stage of cultivation, and a late stage probiotic is used in the late stage of cultivation.
Further, the probiotics in the fertilizer-water period, the probiotics in the growing period and the probiotics in the cultivation later period are added into water containing 0.2-2% of salt and 2-10% of brown sugar by mass percent for activation for 2-6 hours before use.
Preferably, the probiotics in the fertilizer-water period, the probiotics in the growing period and the probiotics in the cultivation later period are added into water containing 1% of salt and 4% of brown sugar by mass for activation for 4 hours before use.
The beneficial effects of the invention are as follows:
the microorganism water quality regulating agent comprehensively uses a plurality of probiotics (algae), improves culture mediums and growth conditions of lactobacillus, rhodopseudomonas palustris, chlorella, saccharomycetes, bacillus subtilis and nitrifying bacteria, establishes different culture and proliferation schemes, can enable each type of microorganism to be put into use after having higher concentration, improves the titer, and ensures the quality and benefit.
The microbial water quality regulator comprises three probiotics, and the probiotics are respectively applicable to the early stage, the middle stage and the later stage of aquaculture by adjusting different types of the probiotics. And (3) corresponding probiotics are added in different aquaculture periods, so that the water body requirements of different aquaculture periods are respectively ensured, a high-quality living environment is provided for aquatic animals, and the product quality is improved. Meanwhile, the probiotics of different types act on the same aquaculture period to realize synergistic interaction, so that the water quality of the aquaculture water body is regulated, a good ecological system is established in the aquaculture water body, the survival rate of aquatic animals is improved, and the aquaculture benefit is increased.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be further clearly and completely described in the following in conjunction with the embodiments of the present invention. It should be noted that the described embodiments are only some embodiments of the present invention, and not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1
A microbial water quality regulator comprises a probiotics agent in a water fertilizer period, a probiotics agent in a growth period and a probiotics agent in a later culture period; the probiotics for the fertilizer-water period comprises 30% of lactic acid bacteria, 40% of chlorella and 30% of saccharomycetes in percentage by mass; the probiotics in the growth period comprises 20% of lactic acid bacteria, 20% of rhodopseudomonas palustris, 30% of saccharomycetes and 30% of bacillus subtilis in percentage by mass; the post-cultivation probiotics comprise, by mass, 20% of lactic acid bacteria, 30% of rhodopseudomonas palustris, 30% of bacillus subtilis and 20% of nitrifying bacteria.
The lactobacillus consists of 60% of lactobacillus fermentation liquor, 20% of shell powder and 20% of freeze-drying protective agent in percentage by mass; the rhodopseudomonas palustris consists of 60% rhodopseudomonas palustris fermentation liquor, 20% shell powder and 20% freeze-drying protective agent in percentage by mass; the chlorella consists of 60% of chlorella fermentation liquor, 20% of shell powder and 20% of freeze-drying protective agent in percentage by mass; the saccharomycete consists of 60% saccharomycete fermentation liquor, 20% shell powder and 20% freeze-drying protective agent in percentage by mass; the bacillus subtilis consists of 60% of bacillus subtilis fermentation liquor, 20% of shell powder and 20% of freeze-drying protective agent in percentage by mass; the nitrifying bacteria consists of 60% nitrifying bacteria fermentation liquor, 20% shell powder and 20% freeze-drying protective agent in percentage by mass.
The lyoprotectant consists of 75% of skimmed milk powder, 5% of lactose and 20% of glycerol in mass percent.
The preparation method of the lactobacillus fermentation broth comprises the following steps:
a1: culturing lactobacillus fermentum, lactobacillus plantarum, lactobacillus casei and streptococcus faecium in a slant solid culture medium at 28-30 ℃ for 36 hours to obtain a first-stage pure culture, then inoculating the first-stage pure culture into a 500mL triangular flask liquid culture medium for 24 hours to obtain a second-stage pure culture, and finally inoculating the second-stage pure culture into a 1.8L mechanical stirring fermentation tank for culturing for 24 hours to obtain lactobacillus fermentation broth, wherein the bacterial count of the lactobacillus fermentation broth is more than or equal to 7log CFU/g; wherein the liquid culture medium is wort culture medium, and the solid culture medium is wort culture medium plus agar.
The culture method of the rhodopseudomonas palustris fermentation liquor comprises the following steps:
b1: preparing a culture medium: 3.5g of sodium acetate, 1.0g of ammonia chloride, 1.2g of monopotassium phosphate, 0.4g of dipotassium phosphate, 0.1g of magnesium chloride, 0.1g of calcium chloride, 0.4g of yeast extract and 0.8g of peptone are weighed, and distilled water is used for constant volume to 1 liter; ph=7.0, autoclaving at 121 ℃ for 20 min;
b2: culturing rhodopseudomonas palustris in a test tube with a prepared culture medium at 28-30 ℃ for 96-144 hours under illumination to obtain a first-stage pure culture, then placing the first-stage pure culture in a 500mL triangular flask for 60-72 hours under constant temperature shaking culture to obtain a second-stage pure culture, and finally inoculating the second-stage pure culture into a 1.8L plastic flask for static culture under illumination at 28-30 ℃ to obtain rhodopseudomonas palustris fermentation broth, wherein the bacterial count of the rhodopseudomonas palustris fermentation broth is more than or equal to 7log CFU/g.
The culture method of the chlorella fermentation broth comprises the following steps:
c1: preparing a culture medium: weighing 0.16g of sodium nitrate, 0.02g of monopotassium phosphate, 0.05g of magnesium sulfate, 0.04g of urea, 0.02g of calcium chloride, 0.02g of sodium bicarbonate, 1.00g of sodium chloride, 4.00mg of ferric citrate, 0.08mg of cupric sulfate pentahydrate, 2.86mg of boric acid, 1.81mg of manganese chloride tetrahydrate and 0.21mg of sodium molybdate dihydrate; the sea water is used for constant volume to 1 liter; ph=7.0, autoclaving at 121 ℃ for 20 min;
c2: culturing chlorella in a test tube filled with a culture medium at 23-24 ℃ for 96-144 hours under illumination to obtain a first-stage pure culture, then culturing in a 500mL triangular flask at constant temperature for 60-72 hours under shaking to obtain a second-stage pure culture, then inoculating to a 1.8L plastic bottle, and culturing at 23-24 ℃ under illumination for static condition to obtain chlorella fermentation broth, wherein the number of algae is more than or equal to 7log CFU/g.
The method for culturing the saccharomycete fermentation liquor comprises the following steps:
d1: preparing a liquid culture medium: weighing 5g of yeast extract, 10g of peptone and 20g of glucose, fixing the volume by distilled water by 1L, adjusting the pH to be 6.4, and autoclaving at 118 ℃ for 20 minutes; preparing a solid culture medium: adding 10g of agar on the basis of the liquid culture medium;
d2: and (3) carrying out slant solid culture on the saccharomyces cerevisiae and the candida utilis at 28-30 ℃ to obtain a first-stage pure culture, then inoculating the first-stage pure culture into a 500mL triangular flask filled with a liquid culture medium, carrying out constant-temperature shaking culture to obtain a second-stage pure culture, and finally inoculating the second-stage pure culture into a 1.8L mechanical stirring fermentation tank to culture for 24 hours to obtain a saccharomycete fermentation broth, wherein the number of bacteria of the saccharomycete fermentation broth is more than or equal to 8log CFU/g.
The method for culturing the bacillus subtilis fermentation broth comprises the following steps:
e1: preparing a culture medium: weighing 10.0g of peptone, 5.0g of yeast extract, 10.0g of sodium chloride and 10.0g of glucose, fixing the volume to 1 liter with distilled water, adjusting the pH to be 7.0, and carrying out damp-heat autoclaving at 118 ℃ for 18 minutes; preparing a solid culture medium: adding 10g of agar on the basis of the liquid culture medium;
e2: performing inclined solid culture on bacillus subtilis at 28-30 ℃ to obtain a first-stage pure culture, then inoculating the first-stage pure culture into a 500mL triangular flask filled with a liquid culture medium, performing constant-temperature shaking culture to obtain a second-stage pure culture, and finally inoculating the second-stage pure culture into a 1.8L mechanical stirring fermentation tank to perform culture for 24 hours to obtain bacillus subtilis fermentation broth, wherein the bacterial count of the bacillus subtilis fermentation broth is more than or equal to 8log CFU/g.
The culture method of the nitrifying bacteria fermentation liquor comprises the following steps:
f1: preparing an ammonia oxidizing bacteria culture medium: weighing 0.5g of ammonium sulfate, 0.3g of sodium chloride, 0.03g of ferrous sulfate, 1.0g of dipotassium hydrogen phosphate, 0.05g of magnesium sulfate, 7.5g of calcium chloride and distilled water to a constant volume of 1 liter; ph=7.0, autoclaving at 121 ℃ for 20 min;
f2: preparing a nitrous acid oxidizing bacteria culture medium: 1.0g of sodium nitrite, 0.03g of magnesium sulfate, 0.01g of manganese sulfate, 0.75g of dipotassium hydrogen phosphate, 1.0g of sodium carbonate, 0.25g of disodium hydrogen phosphate and distilled water with constant volume to 1 liter are weighed; ph=7.0, autoclaving at 121 ℃ for 20 min;
f3: respectively culturing at 28deg.C under 160 r/min for 144 hr to obtain ammonia oxidizing bacteria fermentation broth and nitrite oxidizing bacteria fermentation broth, and mixing to obtain nitrifying bacteria fermentation broth with total bacterial count not less than 8log CFU/g.
Example 2
A microbial water quality regulator comprises a probiotics agent in a water fertilizer period, a probiotics agent in a growth period and a probiotics agent in a later culture period; the probiotics for the fertilizer-water period comprises 20% of lactic acid bacteria, 40% of chlorella and 40% of saccharomycetes in percentage by mass; the probiotics in the growth period comprises 30% of lactic acid bacteria, 10% of rhodopseudomonas palustris, 25% of saccharomycetes and 35% of bacillus subtilis in percentage by mass; the post-cultivation probiotics comprise 20% of lactic acid bacteria, 26% of rhodopseudomonas palustris, 31% of bacillus subtilis and 23% of nitrifying bacteria in percentage by mass.
Wherein the preparation method of lactic acid bacteria, chlorella, yeast, rhodopseudomonas palustris, bacillus subtilis and nitrifying bacteria is the same as that of example 1.
Example 3
A microbial water quality regulator comprises a probiotics agent in a water fertilizer period, a probiotics agent in a growth period and a probiotics agent in a later culture period; the probiotics for the fertilizer-water period comprises 18% of lactic acid bacteria, 46% of chlorella and 36% of saccharomycetes in percentage by mass; the probiotics in the growth period comprises 28% of lactic acid bacteria, 12% of rhodopseudomonas palustris, 24% of saccharomycetes and 36% of bacillus subtilis in percentage by mass; the post-cultivation probiotics comprise 21% of lactic acid bacteria, 25% of rhodopseudomonas palustris, 32% of bacillus subtilis and 22% of nitrifying bacteria by mass percent.
Wherein the preparation method of lactic acid bacteria, chlorella, yeast, rhodopseudomonas palustris, bacillus subtilis and nitrifying bacteria is the same as that of example 1.
Comparative example 1
A microecological preparation for aquaculture (mainly comprising Bacillus subtilis and Bacillus licheniformis) is commercially available.
Comparative example 2
A microbial water quality regulator comprises a probiotics agent in a water fertilizer period, a probiotics agent in a growth period and a probiotics agent in a later culture period; the probiotics for the fertilizer-water period comprises 35% of lactic acid bacteria and 65% of saccharomycetes in percentage by mass; the probiotics in the growth period comprises 28% of lactic acid bacteria, 12% of rhodopseudomonas palustris, 24% of saccharomycetes and 36% of bacillus subtilis in percentage by mass; the post-cultivation probiotics comprises, by mass, 30% of lactic acid bacteria, 30% of rhodopseudomonas palustris and 40% of bacillus subtilis.
Wherein the preparation method of lactobacillus, saccharomycete, rhodopseudomonas palustris and bacillus subtilis is the same as that of example 1.
Comparative example 3
A microbial water quality regulator comprises a probiotics agent in a water fertilizer period, a probiotics agent in a growth period and a probiotics agent in a later culture period; the probiotics for the fertilizer-water period comprises 30% of lactic acid bacteria, 10% of rhodopseudomonas palustris, 25% of saccharomycetes and 35% of bacillus subtilis by mass percent; the probiotic in the growth period comprises 20% of lactic acid bacteria, 40% of chlorella and 40% of saccharomycetes in percentage by mass; the post-cultivation probiotics comprise 20% of lactic acid bacteria, 26% of rhodopseudomonas palustris, 31% of bacillus subtilis and 23% of nitrifying bacteria in percentage by mass.
Wherein the preparation method of lactic acid bacteria, saccharomycetes, rhodopseudomonas palustris, bacillus subtilis, chlorella and nitrifying bacteria is the same as that of example 1.
Comparative example 4
A microbial water quality regulator comprises a probiotics agent in a water fertilizer period, a probiotics agent in a growth period and a probiotics agent in a later culture period; the probiotics for the water-fertilizer period comprises 21% of lactic acid bacteria, 25% of rhodopseudomonas palustris, 32% of bacillus subtilis and 22% of nitrifying bacteria by mass percent; the probiotic in the growth period comprises 18% of lactic acid bacteria, 46% of chlorella and 36% of saccharomycetes in percentage by mass; the post-cultivation probiotics comprise 28% of lactic acid bacteria, 12% of rhodopseudomonas palustris, 24% of saccharomycetes and 36% of bacillus subtilis in percentage by mass.
Wherein the preparation method of lactic acid bacteria, saccharomycetes, rhodopseudomonas palustris, bacillus subtilis, chlorella and nitrifying bacteria is the same as that of example 1.
Detecting data
43 ponds of 2 mu are selected and divided into 7 groups of cultivation comparison test groups, 6 parallel tests are arranged in each group of cultivation tests, and a group of blank control groups are arranged, wherein the blank control groups are free from adding microorganism water quality regulator in the whole process. The microorganism water quality regulators of examples 1 to 3 and comparative examples 1 to 4 are respectively added into the 7-group cultivation comparative test group in the cultivation process, wherein the probiotics in the fertilizer water period, the probiotics in the growth period and the probiotics in the cultivation later period are respectively input in the early stage, the middle stage and the later stage of cultivation, and the comparative example 1 is a commercially available product which is not distinguished from the cultivation period, so that the same microorganism water quality regulator of comparative example 1 is input in the whole cultivation process.
The cultivation method comprises the following steps: the same water source is used in each pond, the water quantity in the ponds is the same, 6 ten thousand shrimp larvae are cultivated in 4 months respectively, and the shrimps are harvested in 7 months. Daily at 9:30 and 18:00 microbial water quality regulator is put in, the total daily feeding amount in the early stage of cultivation is 6-8%, and the total daily feeding amount in the middle and later stages of cultivation is 3-4%. The microbial water quality regulator of each example and comparative example was put in the same manner, in the same amount of time, during the cultivation.
In the invention, the early stage, the middle stage and the later stage of the cultivation are divided according to the individual quantization of the shrimps. Wherein the early stage of cultivation is from the time of raising the seedlings of the shrimps to the time of growing the shrimps to 1000 to 1300 heads per jin; the middle culture period is the early culture period until the shrimps grow to 150-180 per jin; the later culture period is from the middle culture period to the end of culture. For the cultivation process of other aquatic species, the early stage, the middle stage and the later stage of cultivation can be divided according to a conventional distinguishing method in the field.
Pond water quality and transparency were tested separately and the test results are shown in tables 1 to 4. Table 1 shows data of the change of the water quality index with respect to the raw water body at the middle stage of cultivation; table 2 is data of the change of the water quality index with respect to the raw water body at the later stage of cultivation; table 3 is transparency detection data of the water body in the early stage of cultivation; table 4 shows the transparency detection data of the culture water body in the later period of culture. The average of 6 replicates per set of culture trials is shown in tables 1-2.
TABLE 1
TABLE 2
TABLE 3 Table 3
TABLE 4 Table 4
After 7 months of shrimp harvest, weighing 1 jin of shrimps, obtaining the average weight of one shrimp, weighing the total shrimp weight, and calculating the total mantissa of the surviving shrimps by dividing the total shrimp weight by the average weight of the shrimps:
survival (%) = (total mantissa of surviving shrimp/initial mantissa) ×100%.
Disease incidence (%) = area of onset/area of prediction.
The survival rates and disease incidence rates for the different culture test groups are shown in Table 5.
TABLE 5
As can be seen from tables 1 and 2, the microbial water quality regulator according to the embodiment of the invention can maintain stable water quality after different probiotics are added in different cultivation periods, is favorable for intestinal micro-ecology of aquatic animals and inhibits the propagation of pathogenic microorganisms in water, regardless of the change of water quality in mid-cultivation period or late-cultivation period. As can be seen from tables 3 and 4, the probiotics in the water-fertilizer stage are added into the water body in the early stage of cultivation, the water quality is fat and tender, the growth of the prawn larvae is fast, and the transparency of examples 1-3 is lower than that of comparative examples in the early stage of cultivation, which shows that the biomass in the water body is higher after the microbial water quality regulator of the embodiment of the invention is added, and more food is fed to the aquatic animals, thereby being beneficial to the growth of the prawns. In the later period of cultivation, the transparency of the comparative example is obviously lower, and the chemical index is correspondingly higher, which indicates that pathogenic bacteria and blue-green algae in water are propagated, and the health of aquatic animals is easily affected. And after the microbial water quality regulator in examples 1-3 is used, the transparency of the water body in the later period of cultivation is higher. Therefore, the microbial water quality regulator provided by the embodiment of the invention can effectively solve the pollution problem of aquaculture. As is clear from Table 5, the weight gain of the prawns using the microorganism water quality regulator of the invention examples 1 to 3 is remarkable, the feed conversion rate is high, and the yield of the prawns is relatively improved by more than 10% compared with that of the comparative example.
The foregoing examples illustrate only a few embodiments of the invention and are described in detail herein without thereby limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.
Claims (7)
1. The microbial water quality regulator is characterized by comprising a probiotics agent in a water fertilizer period, a probiotics agent in a growth period and a probiotics agent in a later culture period; the probiotics for the fertilizer and water period consists of 10-40% of lactic acid bacteria, 10-50% of chlorella and 10-50% of saccharomycetes in percentage by mass; the probiotics in the growth period consists of, by mass, 10-40% of lactic acid bacteria, 10-40% of rhodopseudomonas palustris, 10-40% of saccharomycetes and 10-40% of bacillus subtilis; the probiotics for the later period of cultivation consists of, by mass, 10-40% of lactic acid bacteria, 10-40% of rhodopseudomonas palustris, 10-40% of bacillus subtilis and 10-40% of nitrifying bacteria;
the lactobacillus adopts lactobacillus fermentum, lactobacillus plantarum, lactobacillus casei and streptococcus faecium; the saccharomycete adopts Saccharomyces cerevisiae and candida utilis; the nitrifying bacteria adopt ammonia oxidizing bacteria and nitrous acid oxidizing bacteria.
2. The microbial water quality regulator according to claim 1, wherein the probiotic agent for the rich water period consists of 30% lactic acid bacteria, 40% chlorella and 30% saccharomycetes in percentage by mass; the probiotics in the growth period consists of 20% of lactic acid bacteria, 20% of rhodopseudomonas palustris, 30% of saccharomycetes and 30% of bacillus subtilis in percentage by mass; the probiotics for the later period of cultivation consists of 20% of lactic acid bacteria, 30% of rhodopseudomonas palustris, 30% of bacillus subtilis and 20% of nitrifying bacteria in percentage by mass.
3. The microbial water quality regulator according to claim 1, wherein the lactic acid bacteria consist of 50-70% of lactic acid bacteria fermentation liquid, 20-40% of shell powder and 10-30% of freeze-drying protective agent in percentage by mass; the rhodopseudomonas palustris consists of 50-70% by mass of rhodopseudomonas palustris fermentation liquor, 20-40% by mass of shell powder and 10-30% by mass of freeze-drying protective agent; the chlorella consists of 50-70% of chlorella fermentation liquor, 20-40% of shell powder and 10-30% of freeze-drying protective agent by mass percent; the saccharomycete consists of 50-70% saccharomycete fermentation liquor, 20-40% shell powder and 10-30% freeze-drying protective agent in percentage by mass; the bacillus subtilis consists of 50-70% of bacillus subtilis fermentation liquor, 20-40% of shell powder and 10-30% of freeze-drying protective agent by mass percent; the nitrifying bacteria consists of 50-70% of nitrifying bacteria fermentation liquor, 20-40% of shell powder and 10-30% of freeze-drying protective agent by mass percent.
4. A microbial water quality conditioner according to claim 3, wherein the lyoprotectant consists of 50% -80% of skimmed milk powder, 2% -20% of lactose and 10% -30% of glycerol by mass percent.
5. The method for preparing the microbial water quality regulator according to any one of claims 1-4, wherein lactobacillus, chlorella and saccharomycetes are mixed according to a proportion to obtain a probiotic agent in a fertilizer-water period; mixing lactobacillus, rhodopseudomonas palustris, saccharomycetes and bacillus subtilis according to a proportion to obtain a probiotic agent in a growth period; and mixing lactobacillus, rhodopseudomonas palustris, bacillus subtilis and nitrifying bacteria in proportion to obtain the probiotic agent in the later period of cultivation.
6. The method for using the microbial water quality regulator according to any one of claims 1 to 4, wherein the probiotic agent is used in a fertilizer water period in a prophase of cultivation, the probiotic agent is used in a growth period in a prophase of cultivation, and the probiotic agent is used in a later period of cultivation.
7. The application method of the microbial water quality regulator according to claim 6, wherein the probiotics in the water fertilizer stage, the probiotics in the growth stage and the probiotics in the later culture stage are added into water containing 0.2-2% of salt and 2-10% of brown sugar by mass percent for activation for 2-6 hours before the probiotics are used.
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