CN108913680A - A kind of microorganism formulation and its application for cultivation water environment ammonia nitrogen degradation - Google Patents
A kind of microorganism formulation and its application for cultivation water environment ammonia nitrogen degradation Download PDFInfo
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- CN108913680A CN108913680A CN201810896519.0A CN201810896519A CN108913680A CN 108913680 A CN108913680 A CN 108913680A CN 201810896519 A CN201810896519 A CN 201810896519A CN 108913680 A CN108913680 A CN 108913680A
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- Prior art keywords
- psb20
- microorganism formulation
- rhodopseudomonas palustris
- microbial inoculum
- ammonia nitrogen
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 44
- 244000005700 microbiome Species 0.000 title claims abstract description 41
- 239000000203 mixture Substances 0.000 title claims abstract description 39
- 238000009472 formulation Methods 0.000 title claims abstract description 37
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 title claims abstract description 24
- 230000015556 catabolic process Effects 0.000 title claims abstract description 16
- 238000006731 degradation reaction Methods 0.000 title claims abstract description 16
- 241000190950 Rhodopseudomonas palustris Species 0.000 claims abstract description 32
- 239000002068 microbial inoculum Substances 0.000 claims abstract description 22
- 230000001580 bacterial effect Effects 0.000 claims abstract description 18
- 238000012258 culturing Methods 0.000 claims abstract description 3
- 241000894006 Bacteria Species 0.000 claims description 37
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 claims description 20
- 238000005286 illumination Methods 0.000 claims description 18
- 239000006071 cream Substances 0.000 claims description 14
- 238000009395 breeding Methods 0.000 claims description 13
- 230000001488 breeding effect Effects 0.000 claims description 13
- 239000007640 basal medium Substances 0.000 claims description 11
- 239000001963 growth medium Substances 0.000 claims description 11
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 claims description 10
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 10
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 10
- 235000019270 ammonium chloride Nutrition 0.000 claims description 10
- 229940041514 candida albicans extract Drugs 0.000 claims description 10
- 239000001632 sodium acetate Substances 0.000 claims description 10
- 235000017281 sodium acetate Nutrition 0.000 claims description 10
- 239000012138 yeast extract Substances 0.000 claims description 10
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 9
- 239000001110 calcium chloride Substances 0.000 claims description 9
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 9
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 9
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 9
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 9
- 238000002360 preparation method Methods 0.000 claims description 9
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical group O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 8
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims description 7
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims description 7
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims description 7
- 239000003223 protective agent Substances 0.000 claims description 7
- 239000007787 solid Substances 0.000 claims description 7
- 229940103272 aluminum potassium sulfate Drugs 0.000 claims description 6
- 238000005189 flocculation Methods 0.000 claims description 6
- 230000016615 flocculation Effects 0.000 claims description 6
- GRLPQNLYRHEGIJ-UHFFFAOYSA-J potassium aluminium sulfate Chemical compound [Al+3].[K+].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O GRLPQNLYRHEGIJ-UHFFFAOYSA-J 0.000 claims description 6
- 239000006228 supernatant Substances 0.000 claims description 6
- 239000011790 ferrous sulphate Substances 0.000 claims description 5
- 235000003891 ferrous sulphate Nutrition 0.000 claims description 5
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 5
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 5
- 229910001629 magnesium chloride Inorganic materials 0.000 claims description 5
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 5
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 5
- 229940099596 manganese sulfate Drugs 0.000 claims description 5
- 239000011702 manganese sulphate Substances 0.000 claims description 5
- 235000007079 manganese sulphate Nutrition 0.000 claims description 5
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 5
- 239000011780 sodium chloride Substances 0.000 claims description 5
- 229910021536 Zeolite Inorganic materials 0.000 claims description 4
- HNPSIPDUKPIQMN-UHFFFAOYSA-N dioxosilane;oxo(oxoalumanyloxy)alumane Chemical compound O=[Si]=O.O=[Al]O[Al]=O HNPSIPDUKPIQMN-UHFFFAOYSA-N 0.000 claims description 4
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 4
- 239000002054 inoculum Substances 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 239000010457 zeolite Substances 0.000 claims description 4
- 239000003795 chemical substances by application Substances 0.000 claims description 3
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims 1
- 239000011575 calcium Substances 0.000 claims 1
- 229910052791 calcium Inorganic materials 0.000 claims 1
- 238000005660 chlorination reaction Methods 0.000 claims 1
- 241000607598 Vibrio Species 0.000 abstract description 9
- 206010047400 Vibrio infections Diseases 0.000 abstract description 9
- 239000005416 organic matter Substances 0.000 abstract description 9
- 238000009360 aquaculture Methods 0.000 abstract description 8
- 244000144974 aquaculture Species 0.000 abstract description 8
- 238000005516 engineering process Methods 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 21
- 230000000694 effects Effects 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 239000000654 additive Substances 0.000 description 3
- 230000000996 additive effect Effects 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000238557 Decapoda Species 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 230000003311 flocculating effect Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 229920003023 plastic Polymers 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 241000238553 Litopenaeus vannamei Species 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-N Nitrous acid Chemical compound ON=O IOVCWXUNBOPUCH-UHFFFAOYSA-N 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-L Phosphate ion(2-) Chemical compound OP([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-L 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- KCIDZIIHRGYJAE-YGFYJFDDSA-L dipotassium;[(2r,3r,4s,5r,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] phosphate Chemical compound [K+].[K+].OC[C@H]1O[C@H](OP([O-])([O-])=O)[C@H](O)[C@@H](O)[C@H]1O KCIDZIIHRGYJAE-YGFYJFDDSA-L 0.000 description 1
- 235000013312 flour Nutrition 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-N phosphoric acid Substances OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 1
- -1 phosphoric acid hydrogen Chemical class 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
Classifications
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- 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
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Inorganic Chemistry (AREA)
- Water Supply & Treatment (AREA)
- Biotechnology (AREA)
- Hydrology & Water Resources (AREA)
- Biodiversity & Conservation Biology (AREA)
- Environmental & Geological Engineering (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention provides a kind of microorganism formulation for cultivation water environment ammonia nitrogen degradation and its applications, belong to aquaculture water quality improving technology field.Microorganism formulation provided by the invention by mass percentage, including 26%~50% Rhodopseudomonas palustris PSB20 microbial inoculum and 50%~74% carrier；The Rhodopseudomonas palustris PSB20 microbial inoculum is obtained by Rhodopseudomonas palustris PSB20 strain culturing；The deposit number of the Rhodopseudomonas palustris PSB20 bacterial strain is CGMCC No:9632.Using microorganism formulation provided by the invention, the ammonia nitrogen and the content of organic matter in aquaculture water environment can significantly reduce, while also can significantly reduce the sum of the vibrios in culture environment of aquatic products, realize the efficient improvement of aquaculture water quality.
Description
Technical field
The invention belongs to aquaculture water quality improving technology fields, and in particular to one kind is used for cultivation water environment ammonia nitrogen degradation
Microorganism formulation and its application.
Background technique
With the development of culture fishery, pond aging year by year, cultivation water, substrate are heavily contaminated, cause to cultivate
Disease frequently occurs, and water quality improvement becomes one of the major measure of healthy aquaculture.In recent years, the research of improver of water quality is not yet
In disconnected development.Traditional improver of water quality is improved using the principle of physics and chemistry by adsorbing or increasing water body dissolved oxygen
Water quality, efficiency is lower, can not solve the requirement of the purification of water quality under current high-density breeding mode.
The Chinese invention patent of Patent No. CN201410517092.0 discloses one kind and changes applied to culture environment of aquatic products
Good microorganism formulation.The microorganism formulation is with Rhodopseudomonas palustris bacterial strain PSB20 and bacillus subtilis strain ND04
Raw material, ammonia nitrogen, nitrous acid in the aquaculture water environment that can degrade, and also there is preferable decomposition to bait residual organic matter
Effect, while the effect for inhibiting growth can also be played to vibrios of curing the disease in breeding water body to a certain extent.But since it is being taken into account
The degradation and maintenance of ammonia nitrogen and nitrite in water body, to ammonia in caused cultivation water environment under high residual bait or other stressed conditions
Degradation effect of meeting an urgent need in the case of nitrogen abruptly increase is relatively limited, so, develop a kind of water environment improvement of energy rapid recovery ammonia nitrogen abruptly increase
Agent be this field practitioner institute there is an urgent need to.
Summary of the invention
In view of this, the purpose of the present invention is to provide one kind for cultivation water environment and the stronger ammonia nitrogen degradation of specificity
Microorganism formulation, ammonia nitrogen and the content of organic matter can be significantly reduced, at the same have the effect of significantly inhibit vibrios.
In order to achieve the above-mentioned object of the invention, the present invention provides following technical scheme：
The present invention provides a kind of microorganism formulations for cultivation water environment ammonia nitrogen degradation, by mass percentage, packet
Include 26%~50% Rhodopseudomonas palustris PSB20 microbial inoculum and 50%~74% carrier；The Rhodopseudomonas palustris
PSB20 microbial inoculum is obtained by Rhodopseudomonas palustris PSB20 strain culturing；The preservation of the Rhodopseudomonas palustris PSB20 bacterial strain
Number be CGMCC No:9632.
Preferably, living bacteria count >=5 × 10 of the Rhodopseudomonas palustris PSB20 microbial inoculum8CFU/g。
Preferably, the carrier is diatomite and/or zeolite powder.
Preferably, the smashing fineness of the carrier is 120~150 mesh.
Preferably, the preparation method of the Rhodopseudomonas palustris PSB20 microbial inoculum includes the following steps：
(1) PSB20 bacterial strain strain is seeded to basal medium, at 25~35 DEG C, illumination cultivation under 2500~3500Lx
72~144h obtains seed liquor；
(2) seed liquor is seeded to expansion breeding culture medium according to the inoculum concentration of volumetric concentration 5%~20%, 30~35
DEG C, 3~5d of illumination cultivation under 2500~3500Lx obtains bacterium solution；
(3) flocculant is added into the bacterium solution, removes supernatant after flocculation, obtains thick bacterium cream；
(4) the thick bacterium cream and protective agent are mixed, obtains Rhodopseudomonas palustris PSB20 microbial inoculum.
Preferably, the basal medium includes the component of following parts by weight：3~4 parts of sodium acetate, potassium dihydrogen phosphate 0.4
~0.8 part, 0.5~1.5 part of ammonium chloride, 0.1~0.5 part of magnesium sulfate, 0.02~0.08 part of calcium chloride, sodium chloride 0.5~1.5
Part, 0.5~1.5 part of yeast extract, 0.01~0.03 part of manganese sulfate, 0.002~0.008 part of ferrous sulfate and 1000 parts of water.
Preferably, described to expand the component that breeding culture medium includes following parts by weight：0.5~1.5 part of ammonium chloride, sodium acetate 2~5
Part, 1~3 part of magnesium chloride, 0.05~0.15 part of calcium chloride, 0.4~0.8 part of potassium dihydrogen phosphate, 0.2~0.6 part of dipotassium hydrogen phosphate,
0.05~0.15 part and 1000 parts of water of yeast extract.
Preferably, the addition volume of the flocculant is the 5~15% of bacterium solution volume, and the flocculant is 0.4~1.0g/
The aluminum potassium sulfate solution of L.
Preferably, it is 0.1~0.5g that the thick bacterium cream of every 1L, which adds protectant amount,；The protective agent is solid trehalose.
The present invention also provides application of the mentioned microorganism preparation in cultivation water environment ammonia nitrogen degradation.
Beneficial effect：The present invention provides a kind of microorganism formulation for cultivation water environment ammonia nitrogen degradation, micro- life
Object preparation includes 26%~50% PSB20 microbial inoculum and 50%~74% carrier.Microorganism formulation provided by the invention is answered
For aquaculture water environment, ammonia nitrogen measured value reduces by 87.5%, and the content of organic matter reduces by 64.4%, and vibrios sum reduces
47.8%.
Specific embodiment
The present invention provides a kind of microorganism formulations for cultivation water environment ammonia nitrogen degradation, by mass percentage, with
The microorganism formulation be 100%, the microorganism formulation include 26%~50% Rhodopseudomonas palustris PSB20 microbial inoculum and
50%~74% carrier.
In the present invention, the Rhodopseudomonas palustris PSB20 microbial inoculum account for the microorganism formulation gross mass 26%~
50%, preferably 30%~48%, more preferably 40%~45%.The present invention is to the Rhodopseudomonas palustris PSB20 microbial inoculum
Source be not particularly limited, expand the red false unit cell in marsh that numerous PSB20 bacterial strain is prepared according to conventional method in that art activation
Bacterium PSB20 microbial inoculum.The PSB20 bacterial strain is Rhodopseudomonas palustris bacterial strain PSB20 (Rhodopseudomonas
Palustris), it was preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on 09 02nd, 2014,
Deposit number is CGMCC No:9632, the patent publication No. for being related to the bacterial strain is CN104498382.
In the present invention, the Rhodopseudomonas palustris PSB20 bacterial preparation process preferably includes following steps：
(1) Rhodopseudomonas palustris PSB20 bacterial strain strain is seeded to basal medium, at 25~35 DEG C, 2500~
72~144h of illumination cultivation under 3500Lx, obtains seed liquor；
(2) seed liquor is seeded to expansion breeding culture medium according to the inoculum concentration of volumetric concentration 5%~20%, 30~35
DEG C, 3~5d of illumination cultivation under 2500~3500Lx obtains bacterium solution；
(3) flocculant is added into the bacterium solution, removes supernatant after flocculation, obtains thick bacterium cream；
(4) protective agent is added into the thick bacterium cream, obtains the Rhodopseudomonas palustris PSB20 microbial inoculum.
Rhodopseudomonas palustris PSB20 bacterial strain strain is first seeded to illumination cultivation in basal medium by the present invention, must be planted
Sub- liquid.In the present invention, the basal medium of this field conventional commercial can be selected in the basal medium.The basal medium
Preferably include the component of following parts by weight：3~4 parts of sodium acetate, 0.4~0.8 part of potassium dihydrogen phosphate, 0.5~1.5 part of ammonium chloride,
0.1~0.5 part of magnesium sulfate, 0.02~0.08 part of calcium chloride, 0.5~1.5 part of sodium chloride, 0.5~1.5 part of yeast extract, manganese sulfate
0.01~0.03 part, 0.002~0.008 part of ferrous sulfate and 1000 parts of water；It more preferably include 3.3 parts of sodium acetate, biphosphate
0.6 part of potassium, 1 part of ammonium chloride, 0.3 part of magnesium sulfate, 0.05 part of calcium chloride, 1 part of sodium chloride, 1 part of yeast extract, 0.023 part of manganese sulfate,
0.005 part and 1000 parts of water of ferrous sulfate.In the present invention, the pH value of the basal medium is preferably 7.5~8.5, more excellent
It is selected as 8.The temperature of the culture is preferably 25~35 DEG C, and more preferably 30 DEG C.The intensity of illumination of the illumination cultivation is preferably
2500~3500Lx, more preferably 3000Lx.The time of the illumination cultivation is preferably 72~144h, more preferably 108h.It is living
Seed liquor is obtained after changing culture；The OD of the seed liquor660Value is 1.4~2.2.
After obtaining seed liquor, the seed liquor is seeded to by the present invention expands illumination cultivation in breeding culture medium, obtains bacterium solution.At this
In invention, the expansion breeding culture medium of this field conventional commercial is may be selected in the breeding culture medium that expands.The expansion breeding culture medium preferably includes
The component of following parts by weight：0.5~1.5 part of ammonium chloride, 2~5 parts of sodium acetate, 1~3 part of magnesium chloride, calcium chloride 0.05~0.15
Part, 0.4~0.8 part of potassium dihydrogen phosphate, 0.2~0.6 part of dipotassium hydrogen phosphate, 0.05~0.15 part of yeast extract and 1000 parts of water；More
Preferably include 1 part of ammonium chloride, 3.5 parts of sodium acetate, 2.0 parts of magnesium chloride, 0.1 part of calcium chloride, 0.6 part of potassium dihydrogen phosphate, phosphoric acid hydrogen
0.4 part of dipotassium, 0.1 part of yeast extract and 1000 parts of water.In the present invention, the pH value for expanding breeding culture medium be preferably 7.0~
7.5, more preferably 7.2.The temperature of the culture is preferably 30~35 DEG C, and more preferably 32 DEG C.The illumination of the illumination cultivation
Intensity is preferably 2500~3500Lx, more preferably 3000~3300Lx.The time of the illumination cultivation is preferably 3~5d, more
Preferably 4d.In the present invention, it is preferable to use daylight illumination for the illumination cultivation, are not available daylight light when meeting cloudy or night
According to when, present invention preferably employs the incandescent lamps of 60W/25L to be irradiated.Bacterium solution is obtained after expanding numerous culture, the bacterium solution is in dark red
Color, OD660≥1.8.In the present invention, the numerous culture of the expansion preferably carries out in transparent plastic agricultural film.
After obtaining bacterium solution, the present invention adds flocculant into the bacterium solution, removes supernatant after flocculation, obtains thick bacterium cream.At this
In invention, the addition volume of the flocculant is preferably the 5~15% of bacterium solution volume, and more preferably 10%.The flocculant is excellent
It is selected as aluminum potassium sulfate solution.The concentration of the aluminum potassium sulfate solution is preferably 0.4~1.0g/L, more preferably 0.5g/L.It is described
Flocculation is preferably carried out in the case where standing environment.The flocculated time is preferably 6~15h, more preferably 8~12h.It is described to remove supernatant
Method preferably be centrifuged, the revolving speed of the centrifugation is preferably 5000rpm, and the time of centrifugation is preferably 15min.
After obtaining thick bacterium cream, the present invention adds protective agent into the thick bacterium cream, obtains Rhodopseudomonas palustris
PSB20 microbial inoculum.In the present invention, it is preferably 0.1~0.5g that the thick bacterium cream of every 1L, which adds protectant amount, more preferably 0.8g.
The protective agent is preferably solid trehalose.The present invention is not particularly limited protectant source, this field routine city
Sell product.
In the present invention, the Rhodopseudomonas palustris PSB20 microbial inoculum living bacteria count preferably >=5 × 108CFU/g,
More preferably >=5 × 109CFU/g.The effect of the Rhodopseudomonas palustris PSB20 microbial inoculum in the present invention is degradation aquaculture
Ammonia nitrogen in water environment decomposes bait residual organic matter, inhibits vibrios growth.
Microorganism formulation provided by the invention further includes carrier.The carrier accounts for the 50% of the microorganism formulation gross mass
~74%, preferably 55%~70%, more preferably 55%~60%.The smashing fineness of the carrier is preferably 120~150
Mesh, more preferably 130~140 mesh.The carrier is preferably diatomite and/or zeolite powder.When the carrier is diatomite and boiling
When the mixture of mountain flour, the mass ratio of diatomite and zeolite powder is preferably 1: 2~2: 1, and more preferably 1.3: 1.6~1.7: 1.4.
In the present invention, the preparation method of mentioned microorganism preparation preferably includes following steps：By the PSB20 microbial inoculum and
It is dry after the carrier mixing to be made.In the present invention, the temperature of the drying is preferably 30~40 DEG C, and more preferably 35 DEG C.
After the drying, microorganism formulation obtained is preferably placed under cryogenic conditions and saves by the present invention.The temperature of the preservation is preferred
It is 2~10 DEG C, more preferably 4 DEG C.
The present invention also provides application of the mentioned microorganism preparation in water environment ammonia nitrogen degradation.By the microorganism formulation
It is watered full pool spilling head after diluting.Per cubic meter of water is preferably 1~5g using the amount of microorganism formulation, more preferably 2g.It is described
Being watered multiple is preferably 5~15 times, and more preferably 10 times.Described splash is preferably spaced 3~10 days, more preferably 7 days.
Below with reference to embodiment to a kind of microorganism formulation applied to cultivation water environment ammonia nitrogen degradation provided by the invention
It is described in detail, but they cannot be interpreted as limiting the scope of the present invention.
Embodiment 1
(1) culture of bacterial strain PSB20：
It takes bacterial strain PSB20 and is seeded to basal medium (sodium acetate 3.3g, potassium dihydrogen phosphate 0.6g, ammonium chloride
1.0g, magnesium sulfate 0.3g, calcium chloride 0.05g, sodium chloride 1.0g, yeast extract 1.0g, manganese sulfate 0.023g, ferrous sulfate
0.005g, water 1000mL, pH 7.5~8.5) in, 27 DEG C, illumination cultivation 72h under 2500Lx, as the OD of culture solution660When up to 1.5
Obtain seed liquor.
Seed liquor is seeded to transparent plastic agricultural film and (includes expansion breeding culture medium by 10% inoculum concentration by volume：Ammonium chloride
1.0g, sodium acetate 3.5g, magnesium chloride 2.0g, calcium chloride 0.1g, potassium dihydrogen phosphate 0.6g, dipotassium hydrogen phosphate 0.4g, yeast extract
0.1g, water 1000mL, pH 7.2), 30~35 DEG C (then using spray cooling when the temperature is excessively high), and utilize sunlight illumination cultivation
3 days (cloudy day and irradiated when night using 60W/25L incandescent lamp) cultivates completion when bacterium solution presents dark red.
(2) concentration and solidification of bacterial strain PSB20：It is 0.5g/ that concentration is added into the bacterium solution that bacterial strain PSB20 has cultivated acquisition
The aluminum potassium sulfate solution of L stands overnight flocculation, removes supernatant；Solid trehalose is added, Rhodopseudomonas palustris PSB20 bacterium is obtained
Agent.Wherein, the additive amount of aluminum potassium sulfate solution is the 5% of the bacterium solution volume that bacterial strain PSB20 has cultivated acquisition；Solid trehalose
Additive amount is：Solid trehalose 0.1g is added in the thick bacterium cream that every 1L is obtained after flocculating.
(3) microorganism formulation is prepared：Diatomite (smashing fineness 120 is added into the bacterium solution for being added to solid trehalose
Mesh), then at 30 DEG C air-dry, packing be stored in 4 DEG C it is spare；The additive amount of diatomite is：Every 1L is obtained dense after flocculating
Diatomite 1000g is added in thick bacterium cream.
Embodiment 2
The preparation-obtained product composition of embodiment 1 is measured：The product with physiological saline prepared by embodiment 1 into
After row gradient dilution, cultivated in 35 DEG C using double-layer agar technique after the coating of each dilution, technical result shows that embodiment 1 is made
PSB20 viable count is 5.82 × 10 in standby product9CFU/mL。
Embodiment 3
Example 1 prepares resulting microorganism formulation, uses on the cultivation white leg Shrimp pool：
Design one group of control group, one group of experimental group；Per cubic meter of water uses 4g embodiment 1 to prepare institute every time in experimental group
Microorganism formulation, when use be watered 10 times of dilution after carry out full pool spilling head, and weekly using primary；Control group is blank pair
According to that is, without using the preparation gained microorganism formulation of embodiment 1；Taken after one month shrimp pool water sample detection its ammonia nitrogen, vibrios sum and
The content of organic matter.
Test result：Control group ammonia nitrogen measured value is 0.8mg/L, vibrios sum is 790CFU/mL, the content of organic matter is
1.6%；And the ammonia nitrogen measured value of experimental group is 0.1mg/L, vibrios sum is 412CFU/mL, the content of organic matter 0.57%.It says
Bright to carry out full pool spilling head using the microorganism formulation prawn provided by the invention pool, ammonia nitrogen, organic matter and the content of vibrios have bright
Aobvious decline.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of microorganism formulation for cultivation water environment ammonia nitrogen degradation, which is characterized in that by mass percentage, including
26%~50% Rhodopseudomonas palustris PSB20 microbial inoculum and 50%~74% carrier；The Rhodopseudomonas palustris PSB20
Microbial inoculum is obtained by Rhodopseudomonas palustris PSB20 strain culturing；The deposit number of the Rhodopseudomonas palustris PSB20 bacterial strain is
CGMCC No:9632。
2. microorganism formulation according to claim 1, which is characterized in that the Rhodopseudomonas palustris PSB20 microbial inoculum
Living bacteria count >=5 × 108CFU/g。
3. microorganism formulation according to claim 1 or 2, which is characterized in that the carrier is diatomite and/or zeolite
Powder.
4. microorganism formulation according to claim 3, the smashing fineness of the carrier is 120~150 mesh.
5. microorganism formulation according to claim 1 or 2, which is characterized in that the Rhodopseudomonas palustris PSB20 microbial inoculum
Preparation method include the following steps：
(1) PSB20 bacterial strain strain is seeded to basal medium, at 25~35 DEG C, illumination cultivation 72 under 2500~3500Lx~
144h obtains seed liquor；
(2) seed liquor is seeded to expansion breeding culture medium according to the inoculum concentration of volumetric concentration 5%~20%, 30~35 DEG C,
3~5d of illumination cultivation under 2500~3500Lx, obtains bacterium solution；
(3) flocculant is added into the bacterium solution, removes supernatant after flocculation, obtains thick bacterium cream；
(4) the thick bacterium cream and protective agent are mixed, obtains Rhodopseudomonas palustris PSB20 microbial inoculum.
6. microorganism formulation according to claim 5, which is characterized in that the basal medium includes following parts by weight
Component：3~4 parts of sodium acetate, 0.4~0.8 part of potassium dihydrogen phosphate, 0.5~1.5 part of ammonium chloride, 0.1~0.5 part of magnesium sulfate, chlorination
0.02~0.08 part of calcium, 0.5~1.5 part of sodium chloride, 0.5~1.5 part of yeast extract, 0.01~0.03 part of manganese sulfate, ferrous sulfate
0.002~0.008 part and 1000 parts of water.
7. microorganism formulation according to claim 5, which is characterized in that the expansion breeding culture medium includes following parts by weight
Component：0.5~1.5 part of ammonium chloride, 2~5 parts of sodium acetate, 1~3 part of magnesium chloride, 0.05~0.15 part of calcium chloride, potassium dihydrogen phosphate
0.4~0.8 part, 0.2~0.6 part of dipotassium hydrogen phosphate, 0.05~0.15 part of yeast extract and 1000 parts of water.
8. according to the described in any item microorganism formulations of claim 5~7, which is characterized in that the addition volume of the flocculant
It is the 5~15% of bacterium solution volume, the flocculant is the aluminum potassium sulfate solution of 0.4~1.0g/L.
9. according to the described in any item microorganism formulations of claim 5~7, which is characterized in that every thick bacterium cream addition protection of 1L
The amount of agent is 0.1~0.5g；The protective agent is solid trehalose.
10. application of any one of claim 1~9 microorganism formulation in cultivation water environment ammonia nitrogen degradation.
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