CN101078004A - Microorganism preparation for modifying water body by using bacterial and manufacturing method thereof - Google Patents
Microorganism preparation for modifying water body by using bacterial and manufacturing method thereof Download PDFInfo
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- CN101078004A CN101078004A CN 200610026796 CN200610026796A CN101078004A CN 101078004 A CN101078004 A CN 101078004A CN 200610026796 CN200610026796 CN 200610026796 CN 200610026796 A CN200610026796 A CN 200610026796A CN 101078004 A CN101078004 A CN 101078004A
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- nitrifier
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- nitrococcus
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 98
- 230000001580 bacterial effect Effects 0.000 title claims description 39
- 238000002360 preparation method Methods 0.000 title claims description 20
- 238000004519 manufacturing process Methods 0.000 title claims description 13
- 244000005700 microbiome Species 0.000 title description 2
- 241000194108 Bacillus licheniformis Species 0.000 claims abstract description 26
- 238000000034 method Methods 0.000 claims abstract description 25
- 241000894006 Bacteria Species 0.000 claims abstract description 22
- 241000108664 Nitrobacteria Species 0.000 claims abstract description 22
- 230000008569 process Effects 0.000 claims abstract description 16
- 238000012216 screening Methods 0.000 claims abstract description 15
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 claims abstract description 13
- 238000000855 fermentation Methods 0.000 claims abstract description 11
- 230000004151 fermentation Effects 0.000 claims abstract description 11
- 238000005516 engineering process Methods 0.000 claims abstract description 10
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 8
- 150000001875 compounds Chemical class 0.000 claims abstract description 3
- 241001495402 Nitrococcus Species 0.000 claims description 46
- 239000000243 solution Substances 0.000 claims description 38
- 230000000694 effects Effects 0.000 claims description 33
- 239000002609 medium Substances 0.000 claims description 28
- 244000061458 Solanum melongena Species 0.000 claims description 23
- 235000002597 Solanum melongena Nutrition 0.000 claims description 23
- 238000012258 culturing Methods 0.000 claims description 23
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 20
- IOVCWXUNBOPUCH-UHFFFAOYSA-N Nitrous acid Chemical compound ON=O IOVCWXUNBOPUCH-UHFFFAOYSA-N 0.000 claims description 20
- 239000000843 powder Substances 0.000 claims description 18
- 239000002068 microbial inoculum Substances 0.000 claims description 16
- 230000001954 sterilising effect Effects 0.000 claims description 15
- 238000004659 sterilization and disinfection Methods 0.000 claims description 15
- 239000006916 nutrient agar Substances 0.000 claims description 13
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 12
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 12
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 12
- 239000003153 chemical reaction reagent Substances 0.000 claims description 12
- 238000002955 isolation Methods 0.000 claims description 12
- 239000007788 liquid Substances 0.000 claims description 12
- 238000002156 mixing Methods 0.000 claims description 12
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 claims description 12
- 230000006872 improvement Effects 0.000 claims description 11
- 238000012360 testing method Methods 0.000 claims description 11
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical class [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 10
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 claims description 10
- 229910019142 PO4 Inorganic materials 0.000 claims description 10
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims description 10
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 10
- 229910021529 ammonia Inorganic materials 0.000 claims description 10
- 239000004202 carbamide Substances 0.000 claims description 10
- 230000000813 microbial effect Effects 0.000 claims description 10
- 229910017604 nitric acid Inorganic materials 0.000 claims description 10
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 10
- 239000010452 phosphate Substances 0.000 claims description 10
- 239000011591 potassium Substances 0.000 claims description 10
- 229910052700 potassium Inorganic materials 0.000 claims description 10
- 229920001817 Agar Polymers 0.000 claims description 8
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 8
- 229910021536 Zeolite Inorganic materials 0.000 claims description 8
- 239000008272 agar Substances 0.000 claims description 8
- 239000001110 calcium chloride Substances 0.000 claims description 8
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 8
- HNPSIPDUKPIQMN-UHFFFAOYSA-N dioxosilane;oxo(oxoalumanyloxy)alumane Chemical compound O=[Si]=O.O=[Al]O[Al]=O HNPSIPDUKPIQMN-UHFFFAOYSA-N 0.000 claims description 8
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 8
- 239000010457 zeolite Substances 0.000 claims description 8
- 239000000463 material Substances 0.000 claims description 7
- 235000015097 nutrients Nutrition 0.000 claims description 7
- 239000010815 organic waste Substances 0.000 claims description 7
- 238000005406 washing Methods 0.000 claims description 7
- 241000726221 Gemma Species 0.000 claims description 6
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 6
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 6
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 claims description 6
- 229940095054 ammoniac Drugs 0.000 claims description 6
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 6
- 229910000397 disodium phosphate Inorganic materials 0.000 claims description 6
- 235000019800 disodium phosphate Nutrition 0.000 claims description 6
- 238000000703 high-speed centrifugation Methods 0.000 claims description 6
- 235000012204 lemonade/lime carbonate Nutrition 0.000 claims description 6
- GVALZJMUIHGIMD-UHFFFAOYSA-H magnesium phosphate Chemical compound [Mg+2].[Mg+2].[Mg+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O GVALZJMUIHGIMD-UHFFFAOYSA-H 0.000 claims description 6
- 229910000400 magnesium phosphate tribasic Inorganic materials 0.000 claims description 6
- 235000013379 molasses Nutrition 0.000 claims description 6
- 238000010298 pulverizing process Methods 0.000 claims description 6
- 238000005070 sampling Methods 0.000 claims description 6
- 210000000582 semen Anatomy 0.000 claims description 6
- 235000010288 sodium nitrite Nutrition 0.000 claims description 6
- 239000002351 wastewater Substances 0.000 claims description 6
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims description 5
- 235000011089 carbon dioxide Nutrition 0.000 claims description 5
- 230000015556 catabolic process Effects 0.000 claims description 5
- 238000006731 degradation reaction Methods 0.000 claims description 5
- -1 nitrite anions Chemical class 0.000 claims description 5
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 claims description 5
- 239000000741 silica gel Substances 0.000 claims description 5
- 229910002027 silica gel Inorganic materials 0.000 claims description 5
- 239000007787 solid Substances 0.000 claims description 5
- 238000000354 decomposition reaction Methods 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 4
- 239000012530 fluid Substances 0.000 claims description 4
- 239000010806 kitchen waste Substances 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- 229940045641 monobasic sodium phosphate Drugs 0.000 claims description 4
- NQPDZGIKBAWPEJ-UHFFFAOYSA-N pentanoic acid group Chemical group C(CCCC)(=O)O NQPDZGIKBAWPEJ-UHFFFAOYSA-N 0.000 claims description 4
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 claims description 4
- 238000003756 stirring Methods 0.000 claims description 4
- 241000396549 Dicraeopsis bacillus Species 0.000 claims description 3
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- 238000010790 dilution Methods 0.000 claims description 3
- 239000012895 dilution Substances 0.000 claims description 3
- 238000011049 filling Methods 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 3
- 238000007689 inspection Methods 0.000 claims description 3
- 241001515939 male-killing Rickettsia from Adalia bipunctata Species 0.000 claims description 3
- YPJKMVATUPSWOH-UHFFFAOYSA-N nitrooxidanyl Chemical group [O][N+]([O-])=O YPJKMVATUPSWOH-UHFFFAOYSA-N 0.000 claims description 3
- 239000002245 particle Substances 0.000 claims description 3
- 235000011152 sodium sulphate Nutrition 0.000 claims description 3
- OTEKOJQFKOIXMU-UHFFFAOYSA-N 1,4-bis(trichloromethyl)benzene Chemical compound ClC(Cl)(Cl)C1=CC=C(C(Cl)(Cl)Cl)C=C1 OTEKOJQFKOIXMU-UHFFFAOYSA-N 0.000 claims description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 2
- 238000012805 post-processing Methods 0.000 claims description 2
- 235000010333 potassium nitrate Nutrition 0.000 claims description 2
- 239000004323 potassium nitrate Substances 0.000 claims description 2
- 239000000047 product Substances 0.000 abstract description 20
- 239000000126 substance Substances 0.000 abstract description 9
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 abstract description 7
- 238000009313 farming Methods 0.000 abstract description 4
- 230000003115 biocidal effect Effects 0.000 abstract description 2
- 239000012263 liquid product Substances 0.000 abstract description 2
- 239000002957 persistent organic pollutant Substances 0.000 abstract description 2
- 230000000295 complement effect Effects 0.000 abstract 1
- 230000029087 digestion Effects 0.000 abstract 1
- 239000003814 drug Substances 0.000 abstract 1
- 239000013586 microbial product Substances 0.000 abstract 1
- 239000003643 water by type Substances 0.000 abstract 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 13
- 238000009360 aquaculture Methods 0.000 description 13
- 244000144974 aquaculture Species 0.000 description 13
- 238000006243 chemical reaction Methods 0.000 description 9
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 8
- 238000009395 breeding Methods 0.000 description 7
- 230000001488 breeding effect Effects 0.000 description 7
- 241000589516 Pseudomonas Species 0.000 description 6
- 230000003647 oxidation Effects 0.000 description 5
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- 241000195493 Cryptophyta Species 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 230000006399 behavior Effects 0.000 description 4
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 4
- 239000012153 distilled water Substances 0.000 description 4
- 230000001976 improved effect Effects 0.000 description 4
- 235000016709 nutrition Nutrition 0.000 description 4
- 230000035764 nutrition Effects 0.000 description 4
- 235000017550 sodium carbonate Nutrition 0.000 description 4
- 229910000029 sodium carbonate Inorganic materials 0.000 description 4
- 241000251468 Actinopterygii Species 0.000 description 3
- MWUXSHHQAYIFBG-UHFFFAOYSA-N Nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- 239000010865 sewage Substances 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 229910002651 NO3 Inorganic materials 0.000 description 2
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 2
- 241000605154 Nitrobacter winogradskyi Species 0.000 description 2
- AZFNGPAYDKGCRB-XCPIVNJJSA-M [(1s,2s)-2-amino-1,2-diphenylethyl]-(4-methylphenyl)sulfonylazanide;chlororuthenium(1+);1-methyl-4-propan-2-ylbenzene Chemical compound [Ru+]Cl.CC(C)C1=CC=C(C)C=C1.C1=CC(C)=CC=C1S(=O)(=O)[N-][C@@H](C=1C=CC=CC=1)[C@@H](N)C1=CC=CC=C1 AZFNGPAYDKGCRB-XCPIVNJJSA-M 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 238000004176 ammonification Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000004172 nitrogen cycle Methods 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 230000003000 nontoxic effect Effects 0.000 description 2
- 235000010289 potassium nitrite Nutrition 0.000 description 2
- 239000004304 potassium nitrite Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- 208000031295 Animal disease Diseases 0.000 description 1
- 241000193755 Bacillus cereus Species 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 description 1
- 241000605159 Nitrobacter Species 0.000 description 1
- 241000605122 Nitrosomonas Species 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 239000005864 Sulphur Substances 0.000 description 1
- IPQVRLSXWJPESU-UHFFFAOYSA-N [N].ON=O Chemical compound [N].ON=O IPQVRLSXWJPESU-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 241001148470 aerobic bacillus Species 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
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- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
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- 239000007789 gas Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 235000015220 hamburgers Nutrition 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 1
- 229910000037 hydrogen sulfide Inorganic materials 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
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- 229910021645 metal ion Inorganic materials 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 239000008239 natural water Substances 0.000 description 1
- 238000006396 nitration reaction Methods 0.000 description 1
- 230000001546 nitrifying effect Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 150000003016 phosphoric acids Chemical class 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
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- 231100000614 poison Toxicity 0.000 description 1
- 230000007096 poisonous effect Effects 0.000 description 1
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- 239000006041 probiotic Substances 0.000 description 1
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Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a Microbial Product which can decompose the organic pollutant in farming waters rapidly and digestion the content of ammonia nitrogen and nitrite in water in time with using nitrobacteria, Bacillus endospores and other bacteria. The characteristic of this product was that the percent by weight before compound was Nitroso-bacteria 20%-40%, nitrifier 20-40%, Bacillus licheniformis 20%-60%. This invention using screening environments and adding concentration technology, at the same time combine directional concentration with fermentation to finish the process of enrich and enlarge the nitrifier by one step . This can reduce the culture time of nitrifier obviously, so that the using of nitrifier would from laboratory to industrialization. The ten times amount about Nitrobacteria than liquid product can complement and assort with high concentration Bacillus endospores to maintain the water with the best farming condition of rich, live, fecund and clear. This product can basically replaced the chemical medicine using in breed aquatics and produce the organic farm produce with green, safe and no antibiotic residue.
Description
Technical field
The present invention relates to a kind of microbial preparation and preparation method who utilizes the strain improvement water body, can be by in water body, directly adding, utilize nitrifier degrade rapidly ammonia nitrogen and nitrous acid content in the water body, simultaneously organic pollution materials such as bait in the pond and animal excrements are decomposed into small-molecule substance, thereby utilized, absorb by useful algae in the water body and other planktonic organism, formation is suitable for the optimum ecotope of aquatic animal to grow, belongs to the agro-ecology product technique field.
Background technology
Flourish along with China aquaculture intensification, cultivation water environment pollutes serious day by day, meanwhile, the discharging of undressed breeding wastewater and industry, sanitary sewage makes water body be subjected to severe contamination, the breeding ecological environment is destroyed, cause the pathogenic micro-organism kind to increase with velocity of propagation and accelerate, the aquaculture organism disease is on the rise, and causes heavy losses for the aquatic products aquaculture.According to incompletely statistics, moderate above breed disease area takes place and accounts for 15%～20% of the breed total area in the annual whole nation, and production loss is above 1,000,000 tons.Because aquatic animal disease that environmental degradation causes or the communicable disease that causes thus increase severely in a large number, become the major obstacle that the restriction culture fishery develops in a healthy way.Life-time service microbiotic and the control of other chemicalses are cultured disease and have been caused a series of environment and social concern, and its outstanding behaviours is microbiotic and the residual increase of harmful chemical in the product, and the fishery products price remains inactive over long periods of time.
At present, the patent of having applied for, application number is CN02156977.0, be entitled as nitrated microbial inoculum of a kind of low temperature and uses thereof, it is characterized in that: a kind of low temperature resistant nitrated microbial inoculum, the nitrated microbial inoculum of this low temperature relates to hamburger bacterium nitrobacter, the Vickers bacterium nitrobacter, bacillus cereus, Bacillus licheniformis, with yeast saccharomyces cerevisiae totally six kinds of bacterial strains, these bacterial strains are after cultivating, consist of low temperature resistant nitrated microbial inoculum in its culture equal volume ratio proportioning, has the effect of removing the ultralow density ammonia nitrogen at a lower temperature, use or adaptive with corresponding bio-reactor separately can be used for the ammonia nitrogen removal of sea farming and batch production circulating seawer high-density breeding and the removal of COD and handles.Nitrifier is only applicable in low temperature breeding environment such as the sea farming in this technology, and range of application is narrow, simultaneously, need cooperate with corresponding bio-reactor, and many, the complex steps of composite bacteria is unfavorable for promoting on producing and using.
Summary of the invention
The purpose of this invention is to provide a kind of microbial preparation that utilizes the strain improvement water body and preparation method who improves the product number of viable.
For realizing above purpose, technical scheme of the present invention provides a kind of microbial preparation that utilizes the strain improvement water body, it is characterized in that the weight percent of described bacterial classification before compound is: nitrococcus 20-40%, nitrifier 20-40%, Bacillus licheniformis 20-60%.
A kind of preparation method who utilizes the microbial preparation of strain improvement water body, it is characterized in that, choose the nitrobacteria of effective degradation of ammonia nitrogen and nitrous acid content, adopt directional technology to carry out enrichment and separate, adopt orienting enriching to combine simultaneously, enrichment, one step of expansion of nitrifier are finished, add genus bacillus simultaneously with decomposition organic waste with zymotechnique, add the carrier that is suitable for nitrifier effect and propagation in post-processing stages, the steps include: the first step: the seed selection of producing bacterial strain
A. bacterium source collection: collect bottom mud in the kitchen waste water, drip two Griess reagents, observe whether solution takes on a red color or pink, judge nitrous acid content according to shade, drip 2 of hexichol ammonia reagents, observe color and whether be blue, judge nitric acid content according to shade;
B. enrichment culture: the darker above-mentioned waste water that gets colors is as the screening source, add in the enrichment medium of nitrifier after the sterilization and nitrococcus in 2% ratio respectively, putting in the 30-38 ℃ of environment 200-240r/min shaking table cultivated 7-10 days, on white plaque, checked the generation of nitrite anions with Griess reagent every several days, cultivate that nutrient solution takes on a red color after 7-10 days, the expression nitrococcus exists, whether there is nitrate radical to exist with the inspection of pentanoic reagent, be blue, nitrifier exists in expression, with shade judgement effect power, select the strong test tube of effect, the nutrient solution that draws in the test tube carries out above step continuation cultivation again, repeat 2-3 time, can constantly eliminate other heterotrophic bacterium, increase Sodium Nitrite or sulfate of ammoniac, obtain nitrifier pregnant solution and nitrococcus pregnant solution respectively to increase the quantity of nitrifier and nitrococcus;
C. strain separating: nitrifier pregnant solution and nitrococcus pregnant solution were fed carbonic acid gas 0.5-1 hour, do on flat board that peptizer prepares and the silica gel plate in the washing agar and to inoculate respectively with enrichment culture liquid, cultivate after 7 days under the 30-38 ℃ of temperature, the bacterium colony of observed needle point size is sublimed bacterial classification, preserves in the test tube of isolation medium;
D. Bacillus licheniformis bacterial classification: adopt soil to utilize nutrient agar to isolate the Bacillus licheniformis bacterial classification, in filling the eggplant bottle of nutrient agar, cultivate and preserve;
E. the production production of hybrid seeds: nitrifier pregnant solution bacterial classification is transferred in the eggplant bottle that the nitrifier isolation medium is housed, nitrococcus pregnant solution bacterial classification is transferred in the eggplant bottle that the nitrococcus isolation medium is housed, the Bacillus licheniformis bacterial classification is transferred in the eggplant bottle of nutrition nutrient agar, under 35-40 ℃ of temperature, cultivated 45-50 hour, treat that eggplant bottle surface lawn is covered with, and can take out when band is yellow in white, put into 2-6 ℃ refrigerator then and preserve;
Second step: the microbial inoculum orienting enriching is cultivated
A. the enrichment culture of nitrococcus: nitrococcus is inoculated in 120 ℃, the nitrococcus enrichment medium of 30min high-temperature sterilization in 2% ratio, mixing speed is 150-200r/min, culture temperature is 30-38 ℃, air flow 1: 1, cultivated 5-7 days, take a sample to check in the culturing process generation situation of nitrous acid is judged concentration effect according to color;
B. the enrichment culture of nitrifier: nitrifier is inoculated in 120 ℃, the nitrifier enrichment medium of 30min high-temperature sterilization in 2% ratio, mixing speed is 150-200r/min, culture temperature is 30-38 ℃, air flow 1: 1, cultivated 5-7 days, take a sample to check in the culturing process generation situation of nitric acid is judged concentration effect according to color;
C. added sulfate of ammoniac and Sodium Nitrite every 3 days in 2% ratio in the above enrichment culture,, survey the quantity that number method (MPN) detects nitrococcus and nitrifier with the dilution cultivation in the culturing process, reaching 10 to improve the concentration effect of nitrococcus and nitrifier
7Finish behind individual/ml to cultivate;
The 3rd step: the independent enlarged culturing of bacterial classification
A. the enlarged culturing of Bacillus licheniformis: eggplant bottle lawn is inoculated in 120-130 ℃, the substratum of 30-45min high-temperature sterilization, mixing speed is 200r/min-240r/min, culture temperature is 35-40 ℃, air flow 1: 1, fermentation time is 22-26 hour, and microscopically is observed when 90-95% gives birth to spore and finished;
B. culture effect detects: the fermented liquid sampling at 80 ℃, 10min heating in water bath, to kill other assorted bacterium, was cultivated 24 hours on nutrient agar plate, check after gemma quantity reaches 2,000,000,000/ml to stop fermentation;
C. the high speed centrifugation of genus bacillus: fermented liquid is high speed centrifugation under the rotating speed of 5000-7000r/min, obtains the lichens gemma spore concentrated solution of high density;
The 4th step: add special carrier
A. microbial inoculum mixes: the weight proportion of nitrococcus, nitrifier, Bacillus licheniformis concentrated solution being pressed 20-40%, 20-40%, 20-60% mixes, and leaves standstill behind the stirring 30-45min;
B. add carrier: with microbial inoculum and carrier with 1: the 3-5 mixed, mixed fluid is granulated on wobbler, carries out fluidized drying again, dried moisture controlled is at 3-5%;
C. crushing screening: behind the dry materials, change between pulverizing and pulverize, temperature of charge is controlled at below 45 ℃; Material after the pulverizing is crossed 50 eye mesh screens, and sieve back coarse fodder is pulverized again; Sampling detects, significant parameter: moisture content≤5%, viable count 2,000,000,000/g; Physical behavior: pale solid particle.
Screening of described nitrifier and enrichment medium are at least two kinds mixture in ammonium sulfate (screening of nitrobacteria and enrichment are alternative with saltpetre), potassium primary phosphate, 7 water magnesium sulfates, 2 water calcium chloride, urea, 7 water SODIUM PHOSPHATE, MONOBASIC, the water, and its weight percent proportioning is ammonium sulfate 0.2-1.0%, potassium primary phosphate 0.02-0.1%, 7 water magnesium sulfate 0.02-0.1%, 2 water calcium chloride 0.02-0.1%, urea 0.1-0.5%, 7 water SODIUM PHOSPHATE, MONOBASIC 0.2-1.0%, water 90-98%.
Described Bacillus licheniformis enlarged culturing base is at least two kinds of mixtures in sterilized water and bean cake powder, urea, ammonium sulfate, Semen Maydis powder, molasses, potassium primary phosphate, Sodium phosphate dibasic, trimagnesium phosphate, the sal epsom.Its weight percent is dregs of beans 2-6%, urea 1-3%, ammonium sulfate 0-1%, Semen Maydis powder 3-10%, molasses 15-20%, potassium primary phosphate 0-1%, Sodium phosphate dibasic 0-1%, trimagnesium phosphate 0-1%, sal epsom 0-1%, sterilized water 56-79%.
The carrier of described interpolation is made up of with lime carbonate oblique native zeolite powder, feed, and its weight percent proportioning is: zeolite powder 60-80%, lime carbonate 20-40%.
The present invention utilizes beneficial microorganism to absorb ammonia nitrogen, nitrous acid nitrogen and hydrogen sulfide etc. in water body, effectively decompose larger molecular organics, suppress a large amount of breedings of pathogenic bacterium simultaneously, pseudomonas, subtilis, nitrobacteria etc. are playing the part of important role to harmful organic substance in the decomposition water body in the physical environment, and they can bring into play oxidation, ammonification.Nitrated, denitrification, separate effects such as sulphur, sulfuration, fixed nitrogen, the movement of animal, remaining bait, the residual body of planktonic organism, chemicals etc. are decomposed into CO2, nitrate, phosphoric acid salt, vitriol etc. rapidly, for the breeding of unit cell algal grown provides nutrition, and the photosynthesis of unit cell algae provides dissolved oxygen for the breathing of organic oxygenolysis and aquaculture organism, constitute a benign ecological circulation, kept and built good condition of water quality.And wherein, can degradation water in ammonia nitrogen and nitrous acid etc. the water body animal is produced mortality harm, have only bacterial classification---the nitrobacteria of wherein a kind of special role.
Nitrobacteria (nitrifying) is a kind of aerobic bacteria, can grow in the water of aerobic or in the layer of sand, and playing the part of very important role in nitrogen cycle purification of water quality process.Belong to the class of property bacterium on one's own account, comprise two kinds of complete different metabolic groups: nitrous acid Pseudomonas (nitrosomonas) and nitric acid Pseudomonas (nitrobacter).Nitrous acid Pseudomonas bacterium is commonly referred to as " the oxidation person of the ammonium ", and because of its unique food source of supporting one's family is an ammonium, the chemical energy that is generated is closed in ammonium and oxidation is enough to make its existence.Ammonium is a kind of ammonia (NH
3) positively charged ion (NH that generates soluble in water
4+), because it just looks like to be a metal ion species in chemically behavior, so called after " ammonium ".The ammonia of gas has pungent stink, and the ammonium of ionic state does not then have special smell, so be easy to be recognized.
When having air to exist, ammonium can be absorbed by nitrous acid Pseudomonas bacterium.They are oxidized to water with its hydrogen atom, replace it with oxygen, so ammonium becomes water and nitrogen oxide soluble in water, latter chemist is called " nitrous acid ", and its reaction formula is as follows: 2NH
3+ 3O
2=2HNO
2+ 2H
2O, this process is called nitrosification.Nitre reaction acid Pseudomonas bacterium is commonly referred to as " the oxidation person of the nitrous acid ", because of its main food source of supporting one's family is a nitrous acid, the chemical energy that is generated is closed in nitrous acid and oxidation is enough to make its existence, and to generate nitric acid be the end product of nitrogen cycle, and its reaction formula is as follows: 2HNO
2+ O
2=2HNO
3, this process is called nitration reaction.Ammonium is oxidized to nitrous acid by nitrobacteria, is oxidized to the reaction of nitric acid subsequently again.Different bacterium carried out by two kinds in this reaction system, must close fit, just do not cause the intermediate NO of reaction
- 2Be detained and accumulate in the water.
Ammonia in the water has two different forms: the one, and ammonia (NH
3), another is ammonium (NH
4+).Ammonia is poisonous, and ammonium is nontoxic.Do not have nitrobacteria to exist in the pond if culture, will inevitably face the danger that ammonia content increases sharply, no matter which kind of method you take can not settle the matter once and for all.Because ammonia is violent in toxicity, as long as water quality deflection alkalescence, a part of ammonium will transform ammonification naturally, when the ammonia concentration in the water reaches the lethal concentration of aquaculture organism, causes major accident loss also just not at all surprising.If but the nitrobacteria that contains sufficient amount in the water is constantly removed ammonium in the water for you, then the stability of the ecological balanced system of whole Chi Shui can obtain to guarantee, and aquaculture organism is grown up in feeding environment safely.
Nitrobacteria in natural water content seldom and its proliferative speed very slow, even under the ideal condition, at least also want one times of time-consuming 24 ~ 36 hours ability propagation.Chief reason is that nitrobacteria need make organism in vivo, they just can't be grown and breed if there are not these organism, making organism then needs the quite time of length, can directly absorb needed organism unlike other different battalion's property bacterium in organic waste.Simultaneously, nitrobacteria is a kind of bacterium of self-operation, and the feature of this bacterium is to dislike organism, if a lot of organic words are arranged, can suppress their growth and breeding on the contrary.Therefore they can't can directly colonize on the organic waste at the bottom of the pond as other different battalion's property bacterium, and must avoid these organic waste, and this can limit the living environment of nitrobacteria virtually.
Number of research projects has been done by research R﹠D institution and universities and colleges about nitrobacteria, under experiment condition, also can produce highly active nitrobacteria, but because nitrobacteria grows on one's own account, reproduction speed is slow, require in the growing environment characteristics such as organic content is low, so the practical application in aquaculture water seldom, simultaneously, the preservation of nitrobacteria also is a technical barrier.At present, nitrifier is only resting on the culture of ornamental fish that water quality is limpid, water body environment is little and stable in the use of degradation of ammonia nitrogen and nitrite, and to using seldom in the large-area aquaculture, result of use is also not obvious.
The art of this patent is from the technology source---bacterial screening is started with, seed selection has nitrated and bacterial classification nitrosification from kitchen waste water, in the laboratory, carry out enrichment and post-directed training, it is low to have changed the nitrifier viable bacteria content, be difficult to produce the drawback of obviously falling ammonia nitrogen and nitrous acid effect, in general nitrifier seed selection, how from sludge sewage, to separate, owing to wherein mostly be trade effluent, so wherein nitrifier and the effect in the aquaculture water differ bigger, and nitrifier content is extremely low in the aquaculture water, generally only 10
2-10
3, separate very difficult.Simultaneously, adopt the method for multistage interpolation reaction substrate to carry out enrichment culture, because nitrobacteria is synthesized self organism by chemical energy, so reproduction speed very slow (conventional next batch is cultivated needs 7-14 days), adopt the method for multistage interpolation reaction to improve greatly concentration effect and time, time can finish in 7 days, and accumulation rate reaches 50%.
Simultaneously, this patent adopts large-scale fermentor tank to carry out industry and cultivates, in cultivation, also adopt the orienting enriching technology, greatly reduce use cost, the subject matter that faces in the growth of nitrifier is exactly that poor growth, accumulation rate are low, production efficiency is the key of cost in the industry growth, and the time shortens 1/2, and efficient just can increase by 1 times.Adopt during the fermentation and add the cultivation effect that the idea culture substrate has orientation, can produce the active bacterium system that similar reaction substrate is had efficient effect.Because the nitrifier individuality in the water body is small, in the procreation process, the tendency that is attached to the fixture appearance is arranged.General liquid preparation can't provide this growing environment, if settle the fixture of some multilist areas to adhere to for it in can pool water within, it be just promptly on the surface attached to these fixtures, and begins to breed.Yet, in the pool water within to settle fixture normally infeasible, reason is that this mode may hinder the movable of fish and is unfavorable for dragging for and catches.This patent has adopted special carrier---the oblique native zeolite powder with organic pollutant in the degraded adsorbed water body, solved this difficult problem dexterously, zeolite powder can adsorb a large amount of active nitrifiers and sporeformer, can provide good propagation and growing environment for nitrifier again.
The as above integrated application of several technology makes the nitrated number of viable of product reach 10
7Individual/more than the ml, be about 10 times of aquarium fish articles for use on the market, to preserve and adopt solid dry powder, validity period is 12 months, and articles for use are liquid product on the market, about actual 3 months usage periods.And price is 1/5 of a like product.Simultaneously, added highly purified Bacillus licheniformis in this patent product, made the product total viable count reach 2,000,000,000/g.Can effectively decompose the organic substance that bait and movement produce in the water body, both interact, improvement water body effect is obvious, the this patent product has been listed national Spark Plan project product in, also be 863 Program project product simultaneously, at present in Shanghai, the big province of aquaculture such as Guangdong, Zhejiang, Jiangsu applies, and obtained good culture benefit and social benefit.
The present invention adopts the nitrated bacterial classification of screening from the special physical environment that is suitable for water body propagation, and in screening, adopted the orienting enriching culture technique, former bacterial classification rate of propagation and cultivation effect have been accelerated greatly, simultaneously, combine with traditional fermentation industry, adopt the orienting enriching technology, the product number of viable has improved 10 times than routine, improve 10 times of quantity, added highly purified Bacillus licheniformis simultaneously, cooperated and decompose oarse-grained organic waste, made both action compensatings, perfect improved effect, adopt carrier can guarantee propagation and the lasting effect of nitrifier, this carrier itself also has the ability of very strong adsorbed water body impurity simultaneously, adopts the solid particulate packing, usage period prolongs more than 4 times, makes the use of nitrifier move towards cultivation site from the laboratory.
The invention has the advantages that:
1. adopt unique screening environment and enrichment adding technique, guaranteed the high efficiency and the high yield of original strain;
2. the technology that adopts the orienting enriching of original creation to combine with zymotechnique with the enrichment of nitrifier, enlarge one and go on foot and finish, has significantly reduced the incubation time of nitrifier;
3. the Bacillus licheniformis of product middle and high concentration is in time adsorbed decomposition with larger molecular organicses such as water-bed bait, movements, nitrifier has guaranteed that ammonia nitrogen and the nitrite in the water body in time is decomposed into nontoxic nitrate, for useful algae and planktonic organism in the water provide nutrition, both effects one in front and one in back, replenish mutually, guarantee that water body environment maintains the best breed state of " fertile, alive, tender, refreshing ";
4. be suitable for adsorbing the characteristics of growth according to nitrifier, both added and to have absorbed organic waste in the water body, good growth of nitrifier and propagation environment can be provided again, improved survival time and the action time of nitrifier greatly, compare conventional products, quantitative value improves more than 10 times;
5. the present invention adopts the probiotics of occurring in nature seed selection, meets feedstuff industry 318 bacterial classifications and adds standard, can significantly reduce and alternative feed in antibiotic use, produce green, safety, the residual organic farm products of antibiotic-free;
6. adopt food crop and common feedstuffs carrier as the main medium raw material, reduced production cost, use fermentation and production unit commonly used simultaneously, be easy in aquaculture, promote.
Embodiment
The invention will be further described below in conjunction with embodiment.
Embodiment 1
The first step: the seed selection of producing bacterial strain
A. bacterium source collection: collect in the waste water bottom mud a little and waste water altogether about 300ml at the kitchen waste water sewage draining exit, drips two Griess reagents, observe whether solution takes on a red color or pink, according to shade judgement nitrous acid content.Drip 2 of pentanoic reagent, observe color and whether be blue, judge nitric acid content according to shade;
B. enrichment culture: the darker above-mentioned waste water that gets colors is as the screening source, add in the enrichment medium of nitrifier after the sterilization and nitrococcus in 2% ratio respectively, putting in 30 ℃ of environment the 200r/min shaking table cultivated 7 days, on white plaque, checked the generation of nitrite anions every 2 days with Griess reagent, cultivate
Nutrient solution takes on a red color after 7 days, whether the expression nitrococcus exists, have nitrate radical to exist with the inspection of pentanoic reagent, is blue, nitrifier exists in expression, with shade judgement effect power by force, select the strong test tube of effect, draw nutrient solution in the test tube and carry out above step again and continue to cultivate, repeat 2-3 time, can constantly eliminate other heterotrophic bacterium, increase Sodium Nitrite or sulfate of ammoniac, obtain nitrifier pregnant solution and nitrococcus pregnant solution respectively to increase the quantity of nitrifier and nitrococcus;
The nitrifier isolation medium is: ammonium sulfate 0.5%, dipotassium hydrogen phosphate 0.07%, 7 water magnesium sulfates 0.05%, 2 water calcium chloride 0.05%, distilled water 99.33%, transfer to pH7.8 with 5% yellow soda ash, and add 15% washing agar again and make separating plate;
The nitrite bacteria enrichment medium: potassium nitrite 0.5%, dipotassium hydrogen phosphate 0.07%, 7 water magnesium sulfates 0.05%, 2 water calcium chloride 0.05%, distilled water 99.33%, transfer pH7.8,121 ℃, 30min autoclaving with 5% yellow soda ash;
C. strain separating: nitrifier pregnant solution and nitrococcus pregnant solution were fed carbonic acid gas 0.5 hour, do on flat board that peptizer prepares and the silica gel plate in the washing agar and to inoculate respectively with enrichment culture liquid, cultivate after 7 days for 30 ℃, the bacterium colony of observed needle point size is sublimed bacterial classification, preserves in the test tube of isolation medium respectively;
The nitrifier isolation medium is: ammonium sulfate 0.5%, dipotassium hydrogen phosphate 0.07%, 7 water magnesium sulfates 0.05%, 2 water calcium chloride, 0.05%, distilled water 99.33%, transfer to pH7.8 with 5% yellow soda ash, and add 15% washing agar again and make separating plate;
Nitrite bacteria concentration and separation substratum is: potassium nitrite 0.5%, dipotassium hydrogen phosphate 0.07%, 7 water magnesium sulfates 0.05%, 2 water calcium chloride, 0.05%, distilled water 99.33%, transfer to pH7.8 with 5% yellow soda ash, add 15% washing agar again and make separating plate;
D. Bacillus licheniformis bacterial classification: the Bacillus licheniformis bacterial classification Bacillus licheniformis (13123) that utilizes nutrient agar to be separated in soil, cultivate in filling the eggplant bottle of LB and preserve;
Nutrient agar is: extractum carnis 3g, peptone 10g, NaCl5g, agar 15-20g, water 1000ml, pH7.0-7.2;
E. the production production of hybrid seeds: nitrifier pregnant solution bacterial classification is transferred in the eggplant bottle that the nitrifier substratum is housed, nitrococcus pregnant solution bacterial classification is transferred in the eggplant bottle that the nitrococcus substratum is housed, the Bacillus licheniformis bacterial classification is transferred in the eggplant bottle that nutrient agar is housed, under 35-40 ℃ of temperature, cultivated 45-50 hour, treat that eggplant bottle surface lawn is covered with, and can take out when band is yellow in white, put into 2-6 ℃ refrigerator then and preserve;
Second step: the microbial inoculum orienting enriching is cultivated
A. the enrichment culture of nitrococcus: nitrococcus is inoculated in 120 ℃, the nitrococcus enrichment medium of 30min high-temperature sterilization in 2% ratio, mixing speed is 200r/min, culture temperature is 30 ℃, air flow 1: 1, cultivated 7 days, take a sample to check in the culturing process generation situation of nitrous acid is judged concentration effect according to color;
B. the enrichment culture of nitrifier: nitrifier is inoculated in 120 ℃, the nitrifier enrichment medium of 30min high-temperature sterilization in 2% ratio, mixing speed is 200r/min, culture temperature is 30 ℃, air flow 1: 1, cultivated 5-7 days, take a sample to check in the culturing process generation situation of nitric acid is judged concentration effect according to color;
C. added sulfate of ammoniac and Sodium Nitrite every 3 days in 2% ratio in the above enrichment culture, to improve the concentration effect of nitrobacteria;
D. concentration effect check: cultivate the quantity that survey number method (MPN) detects nitrococcus and nitrifier with dilution in the culturing process, reaching 10
7Finish behind individual/ml to cultivate;
The 3rd step: the independent enlarged culturing of bacterial classification
A. the enlarged culturing of Bacillus licheniformis: eggplant bottle lawn is inoculated in 121 ℃, the substratum of 30min high-temperature sterilization, mixing speed is 200r/min, and culture temperature is 35 ℃, air flow 1: 1, fermentation time is 26 hours, and microscopically is observed when 90-95% gives birth to spore and finished;
B. culture effect detects: the fermented liquid sampling at 80 ℃, 10min heating in water bath, to kill other assorted bacterium, is cultivated 24hour on nutrient agar plate, check after gemma quantity reaches 2,000,000,000/ml to stop fermentation;
C. the high speed centrifugation of genus bacillus: fermented liquid is high speed centrifugation under the rotating speed of 5000-7000r/min, obtains the lichens gemma spore concentrated solution of high density;
Genus bacillus enlarged culturing base: bean cake powder 2%, urea 1%, ammonium sulfate 0.5%, Semen Maydis powder 3%, molasses 15%, potassium primary phosphate 0.5%, Sodium phosphate dibasic 0.5%, trimagnesium phosphate 0.5%, sal epsom 0.5%, sterilized water 76.5%;
The 4th step: add special carrier
A. microbial inoculum mixes: nitrococcus, nitrifier, Bacillus licheniformis concentrated solution are mixed by 40%, 40%, 20% quality proportioning, leave standstill behind the stirring 45min;
B. add carrier: with microbial inoculum and carrier with 1: 5 mixed, carrier is selected the oblique native zeolite powder of 80 purposes, 80% weight and the feed lime carbonate of 20% weight for use, mixed fluid carries out fluidized drying again with granulating on the wobbler, and dried moisture controlled is at 3-5%;
C. crushing screening: behind the dry materials, change between pulverizing and pulverize, temperature of charge is controlled at below 45 ℃.Material after the pulverizing is crossed 50 eye mesh screens, and sieve back coarse fodder is pulverized again;
D. notify the sampling of QC department to detect significant parameter: moisture content≤5%, viable count 2,000,000,000/g; Physical behavior: pale solid particle.
Embodiment 2
The first step: the seed selection of producing bacterial strain
C. strain separating: before separating pregnant solution was fed carbonic acid gas 0.5 hour, do on flat board that peptizer prepares and the silica gel plate in the washing agar and to inoculate respectively with enrichment culture liquid, cultivate after 7 days under 30 ℃ of temperature, the bacterium colony of observed needle point size is sublimed bacterial classification, preserves in the test tube of isolation medium.
E. the production production of hybrid seeds: nitrifier pregnant solution bacterial classification is transferred in the eggplant bottle that the nitrifier isolation medium is housed, nitrococcus pregnant solution bacterial classification is transferred in the eggplant bottle that the nitrococcus isolation medium is housed, the Bacillus licheniformis kind is transferred in the eggplant bottle of nutrition nutrient agar, under 35 ℃ of temperature, cultivated 45 hours, band eggplant bottle surface lawn is covered with, and can take out when band is yellow in white, put into 2-6 ℃ refrigerator then and preserve;
Nitrifier enrichment medium: (NH
4) SO
4(or KNO
3) 0.7%, KH
2PO
40.05%, MgSO
47H
2O0.08%, CaCl
22H
2O, 0.07%, Na
2HPO
40.05%, H
2O 98%, uses 5%NaCO
3Transfer pH8.0.121 ℃, 30min autoclaving; Nutrient solution is added with 2% ratio and the making silica gel plate.
Other step is with embodiment 1;
Second step: the microbial inoculum orienting enriching is cultivated
A. the enrichment culture of nitrococcus: nitrococcus is inoculated in 121 ℃, the nitrococcus enrichment medium of 30min high-temperature sterilization in 2% ratio, mixing speed is 200r/min, culture temperature is 30 ℃, air flow 1: 0.8, cultivated 7 days, take a sample to check in the culturing process generation situation of nitrous acid is judged concentration effect according to color;
B. the enrichment culture of nitrifier: nitrifier is inoculated in 120 ℃, the nitrifier enrichment medium of 30min high-temperature sterilization in 2% ratio, mixing speed is 200r/min, culture temperature is 30 ℃, air flow 1: 0.8, cultivated 7 days, take a sample to check in the culturing process generation situation of nitric acid is judged concentration effect according to color.
All the other steps are with example 1.
The 3rd step: the independent enlarged culturing of bacterial classification
A. the enlarged culturing of Bacillus licheniformis: eggplant bottle lawn is inoculated in 121 ℃, the substratum of 30min high-temperature sterilization, mixing speed is 200r/min, and culture temperature is 35 ℃, air flow 1: 0.8, fermentation time is 26 hours, and microscopically is observed when 90-95% gives birth to spore and finished.；
Bacillus licheniformis enlarged culturing base is: its weight percent is dregs of beans 6%, urea 3%, ammonium sulfate 1%, Semen Maydis powder 8%, molasses 20%, potassium primary phosphate 1%, Sodium phosphate dibasic 1%, trimagnesium phosphate 1%, sal epsom 1%, sterilized water 58%.
All the other are with embodiment 1;
The 4th step: add special carrier
A. microbial inoculum mixes: nitrococcus, nitrifier, Bacillus licheniformis concentrated solution liquid are mixed by 35%, 40%, 25% weight proportion, leave standstill behind the stirring 45min.
B. add carrier: with 1: 4 mixed, carrier selected for use 60 order zeolite powders 60%, lime carbonate 40% mixed fluid to granulate on wobbler, carries out fluidized drying again with microbial inoculum and carrier, and dried moisture controlled is at 3-5%.
All the other are with embodiment 1.
Culture the pool by per 10 mu during use, 1 meter depth of water, this product 1kg that directly splashes uses that ammonia nitrogen and nitrite content will obviously descend in the aquaculture water of back, in the use experiment at aquatic products station, area, Qingpu, Shanghai, the ammonia-nitrogen content average specific control group on the test pool has reduced by 72.2%, nitrite has reduced by 60%, and dissolved oxygen has improved 9.9%, and water quality will become grass green by muddiness, 250/ml of harmful algae quantity decline, water body are in the best of " fertile, alive, tender, refreshing " and culture state.
Claims (5)
1. a microbial preparation that utilizes the strain improvement water body is characterized in that, the weight percent of described bacterial classification before compound is: nitrococcus 20-40%, nitrifier 20-40%, Bacillus licheniformis 20-60%.
2. a kind of preparation method who utilizes the microbial preparation of strain improvement water body according to claim 1, it is characterized in that, choose the nitrobacteria of effective degradation of ammonia nitrogen and nitrous acid content, adopt directional technology to carry out enrichment and separate, adopt orienting enriching to combine simultaneously, enrichment, one step of expansion of nitrifier are finished, add genus bacillus simultaneously with decomposition organic waste with zymotechnique, add the carrier that is suitable for nitrifier effect and propagation in post-processing stages, the steps include:
The first step: the seed selection of producing bacterial strain
A. bacterium source collection: collect bottom mud in the kitchen waste water, drip two Griess reagents, observe whether solution takes on a red color or pink, judge nitrous acid content according to shade, drip 2 of hexichol ammonia reagents, observe color and whether be blue, judge nitric acid content according to shade;
B. enrichment culture: the darker above-mentioned waste water that gets colors is as the screening source, add in the enrichment medium of nitrifier after the sterilization and nitrococcus in 2% ratio respectively, putting in the 30-38 ℃ of environment 200-240r/min shaking table cultivated 7-10 days, on white plaque, checked the generation of nitrite anions with Griess reagent every 2-3 days, cultivate that nutrient solution takes on a red color after 7-10 days, the expression nitrococcus exists, whether there is nitrate radical to exist with the inspection of pentanoic reagent, be blue, nitrifier exists in expression, with shade judgement effect power, select the strong test tube of effect, the nutrient solution that draws in the test tube carries out above step continuation cultivation again, repeat 2-3 time, can constantly eliminate other heterotrophic bacterium, increase Sodium Nitrite or sulfate of ammoniac, obtain nitrifier pregnant solution and nitrococcus pregnant solution respectively to increase the quantity of nitrifier and nitrococcus;
C. strain separating: nitrifier pregnant solution and nitrococcus pregnant solution were fed carbonic acid gas 0.5-1 hour, do on flat board that peptizer prepares and the silica gel plate in the washing agar and to inoculate respectively with enrichment culture liquid, cultivate after 7 days under the 30-38 ℃ of temperature, the bacterium colony of observed needle point size is sublimed bacterial classification, preserves in the test tube of isolation medium;
D. Bacillus licheniformis bacterial classification: utilize nutrient agar to isolate the Bacillus licheniformis bacterial classification, in filling the eggplant bottle of nutrient agar, cultivate and preserve;
E. the production production of hybrid seeds: nitrifier pregnant solution bacterial classification is transferred in the eggplant bottle that the nitrifier isolation medium is housed, nitrococcus pregnant solution bacterial classification is transferred in the eggplant bottle that the nitrococcus isolation medium is housed, the Bacillus licheniformis bacterial classification is transferred in the eggplant bottle that nutrient agar is housed, under 35-40 ℃ of temperature, cultivated 45-50 hour, treat that eggplant bottle surface lawn is covered with, and can take out when band is yellow in white, put into 2-6 ℃ refrigerator then and preserve;
Second step: the microbial inoculum orienting enriching is cultivated
A. the enrichment culture of nitrococcus: nitrococcus is inoculated in 120 ℃, the nitrococcus enrichment medium of 30min high-temperature sterilization in 2% ratio, mixing speed is 150-200r/min, culture temperature is 30-38 ℃, air flow 1: 1, cultivated 5-7 days, take a sample to check in the culturing process generation situation of nitrous acid is judged concentration effect according to color;
B. the enrichment culture of nitrifier: nitrifier is inoculated in 120 ℃, the nitrifier enrichment medium of 30min high-temperature sterilization in 2% ratio, mixing speed is 150-200r/min, culture temperature is 30-38 ℃, air flow 1: 1, cultivated 5-7 days, take a sample to check in the culturing process generation situation of nitric acid is judged concentration effect according to color;
C. added sulfate of ammoniac and Sodium Nitrite every 3 days in 2% ratio in the above enrichment culture,, survey the quantity that number method (MPN) detects nitrococcus and nitrifier with the dilution cultivation in the culturing process, reaching 10 to improve the concentration effect of nitrococcus and nitrifier
7Finish behind individual/ml to cultivate;
The 3rd step: the independent enlarged culturing of bacterial classification
A. the enlarged culturing of Bacillus licheniformis: eggplant bottle lawn is inoculated in sub-120-130 ℃, the substratum of 30-45min high-temperature sterilization, mixing speed is 200r/min-240r/min, culture temperature is 35-40 ℃, air flow 1: 1, fermentation time is 22-26 hour, and microscopically is observed when 90-95% gives birth to spore and finished;
B. culture effect detects: the fermented liquid sampling at 80 ℃, 10min heating in water bath, to kill other assorted bacterium, was cultivated 24 hours on nutrient agar plate, check after gemma quantity reaches 2,000,000,000/ml to stop fermentation;
C. the high speed centrifugation of genus bacillus: fermented liquid is high speed centrifugation under the rotating speed of 5000-7000r/min, obtains the lichens gemma spore concentrated solution of high density;
The 4th step: add special carrier
A. microbial inoculum mixes: the weight proportion of nitrococcus, nitrifier, Bacillus licheniformis concentrated solution being pressed 20-40%, 20-40%, 20-60% mixes, and leaves standstill behind the stirring 30-45min;
B. add carrier: with microbial inoculum and carrier with 1: the 3-5 mixed, mixed fluid is granulated on wobbler, carries out fluidized drying again, dried moisture controlled is at 3-5%;
C. crushing screening: behind the dry materials, change between pulverizing and pulverize, temperature of charge is controlled at below 45 ℃, and the material after the pulverizing is crossed 50 eye mesh screens, and sieve back coarse fodder is pulverized again; Sampling detects, significant parameter: moisture content≤5%, viable count 2,000,000,000/g; Physical behavior: pale solid particle.
3. a kind of preparation method who utilizes the microbial preparation of strain improvement water body according to claim 2, it is characterized in that, described nitrifier screening and enrichment medium are ammonium sulfate (screening of nitrobacteria and enrichment substitute with saltpetre), potassium primary phosphate, 7 water magnesium sulfates, 2 water calcium chloride, urea, 7 water SODIUM PHOSPHATE, MONOBASIC, at least two kinds mixture in the water, its weight percent proportioning is ammonium sulfate 0.2-1.0%, potassium primary phosphate 0.02-0.1%, 7 water magnesium sulfate 0.02-0.1%, 2 water calcium chloride 0.02-0.1%, urea 0.1-0.5%, 7 water SODIUM PHOSPHATE, MONOBASIC 0.2-1.0%, water 90-98%.
4. a kind of preparation method who utilizes the microbial preparation of strain improvement water body according to claim 2, it is characterized in that described Bacillus licheniformis enlarged culturing base is at least two kinds of mixtures in sterilized water and bean cake powder, urea, ammonium sulfate, Semen Maydis powder, molasses, potassium primary phosphate, Sodium phosphate dibasic, trimagnesium phosphate, the sal epsom.Its weight percent is dregs of beans 2-6%, urea 1-3%, ammonium sulfate 0-1%, Semen Maydis powder 3-10%, molasses 15-20%, potassium primary phosphate 0-1%, Sodium phosphate dibasic 0-1%, trimagnesium phosphate 0-1%, sal epsom 0-1%, sterilized water 56-79%.
5. a kind of preparation method who utilizes the microbial preparation of strain improvement water body according to claim 2, it is characterized in that, the carrier of described interpolation is made up of with lime carbonate oblique native zeolite powder, feed, and its weight percent proportioning is: zeolite powder 60-80%, lime carbonate 20-40%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200610026796 CN101078004A (en) | 2006-05-23 | 2006-05-23 | Microorganism preparation for modifying water body by using bacterial and manufacturing method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN 200610026796 CN101078004A (en) | 2006-05-23 | 2006-05-23 | Microorganism preparation for modifying water body by using bacterial and manufacturing method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101402934B (en) * | 2008-10-30 | 2011-07-20 | 河南省龙腾高科实业有限公司 | Water quality purification bacterium agent from mix fermentation production with multiple bacterials and production method thereof |
| CN102286390A (en) * | 2010-06-18 | 2011-12-21 | 辽宁北方环境保护有限公司 | Method for screening low temperature resisting nitrifying bacteria |
| CN102329005A (en) * | 2011-06-20 | 2012-01-25 | 大连金砣水产食品有限公司 | Microbial preparation for aquaculture and preparation method thereof |
| CN103396966A (en) * | 2013-08-09 | 2013-11-20 | 湖北科亮生物环保科技有限公司 | Method for producing high-efficiency composite autotrophic nitrifying bacteria |
| CN103395890A (en) * | 2013-07-19 | 2013-11-20 | 上海创博生态工程有限公司 | Microbial preparation for improving freshwater aquaculture water body and preparation method thereof |
| CN105211104A (en) * | 2015-08-28 | 2016-01-06 | 上海创博生态工程有限公司 | A kind of preparation method of pet breeding environment modifying agent |
| CN106047772A (en) * | 2016-08-09 | 2016-10-26 | 湖北凌卓生物工程有限公司 | Composite microbial agent for treating high-ammonia-nitrogen sewage and preparation method and application thereof |
| CN118581017A (en) * | 2024-08-06 | 2024-09-03 | 四川水王子环境科技有限公司 | A composite biological agent for circulating cooling water and its preparation method and application |
| CN119265089A (en) * | 2024-12-12 | 2025-01-07 | 四川清和科技有限公司 | Bacillus licheniformis, polluted water body repair agent and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101402934B (en) * | 2008-10-30 | 2011-07-20 | 河南省龙腾高科实业有限公司 | Water quality purification bacterium agent from mix fermentation production with multiple bacterials and production method thereof |
| CN102286390A (en) * | 2010-06-18 | 2011-12-21 | 辽宁北方环境保护有限公司 | Method for screening low temperature resisting nitrifying bacteria |
| CN102329005A (en) * | 2011-06-20 | 2012-01-25 | 大连金砣水产食品有限公司 | Microbial preparation for aquaculture and preparation method thereof |
| CN102329005B (en) * | 2011-06-20 | 2013-04-24 | 大连金砣水产食品有限公司 | Preparation method of microbial preparation for aquaculture |
| CN103395890A (en) * | 2013-07-19 | 2013-11-20 | 上海创博生态工程有限公司 | Microbial preparation for improving freshwater aquaculture water body and preparation method thereof |
| CN103395890B (en) * | 2013-07-19 | 2015-06-03 | 上海创博生态工程有限公司 | Microbial preparation for improving freshwater aquaculture water body and preparation method thereof |
| CN103396966A (en) * | 2013-08-09 | 2013-11-20 | 湖北科亮生物环保科技有限公司 | Method for producing high-efficiency composite autotrophic nitrifying bacteria |
| CN105211104A (en) * | 2015-08-28 | 2016-01-06 | 上海创博生态工程有限公司 | A kind of preparation method of pet breeding environment modifying agent |
| CN106047772A (en) * | 2016-08-09 | 2016-10-26 | 湖北凌卓生物工程有限公司 | Composite microbial agent for treating high-ammonia-nitrogen sewage and preparation method and application thereof |
| CN118581017A (en) * | 2024-08-06 | 2024-09-03 | 四川水王子环境科技有限公司 | A composite biological agent for circulating cooling water and its preparation method and application |
| CN119265089A (en) * | 2024-12-12 | 2025-01-07 | 四川清和科技有限公司 | Bacillus licheniformis, polluted water body repair agent and application thereof |
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