CN109897804A - One plant has Zhuo Beier Salmonella and its application of nitrification and denitrification function simultaneously - Google Patents
One plant has Zhuo Beier Salmonella and its application of nitrification and denitrification function simultaneously Download PDFInfo
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- CN109897804A CN109897804A CN201910234181.7A CN201910234181A CN109897804A CN 109897804 A CN109897804 A CN 109897804A CN 201910234181 A CN201910234181 A CN 201910234181A CN 109897804 A CN109897804 A CN 109897804A
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- 241000607142 Salmonella Species 0.000 title claims abstract description 36
- 241000196324 Embryophyta Species 0.000 title abstract description 12
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims abstract description 96
- 229910052757 nitrogen Inorganic materials 0.000 claims abstract description 48
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 36
- 241000894006 Bacteria Species 0.000 claims abstract description 22
- MMDJDBSEMBIJBB-UHFFFAOYSA-N [O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O.[NH6+3] Chemical compound [O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O.[NH6+3] MMDJDBSEMBIJBB-UHFFFAOYSA-N 0.000 claims abstract description 22
- JVMRPSJZNHXORP-UHFFFAOYSA-N ON=O.ON=O.ON=O.N Chemical compound ON=O.ON=O.ON=O.N JVMRPSJZNHXORP-UHFFFAOYSA-N 0.000 claims abstract description 20
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 claims abstract description 20
- 241001569440 Zobellella sp. Species 0.000 claims abstract description 10
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 18
- 239000001963 growth medium Substances 0.000 claims description 9
- 229910021529 ammonia Inorganic materials 0.000 claims description 5
- 239000003795 chemical substances by application Substances 0.000 claims description 5
- 239000000470 constituent Substances 0.000 claims description 2
- GQPLMRYTRLFLPF-UHFFFAOYSA-N nitrous oxide Inorganic materials [O-][N+]#N GQPLMRYTRLFLPF-UHFFFAOYSA-N 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims description 2
- 235000015170 shellfish Nutrition 0.000 claims description 2
- 238000000034 method Methods 0.000 abstract description 16
- 230000008569 process Effects 0.000 abstract description 10
- 230000000694 effects Effects 0.000 abstract description 9
- 238000009825 accumulation Methods 0.000 abstract description 7
- 150000001875 compounds Chemical class 0.000 abstract description 5
- 230000006872 improvement Effects 0.000 abstract description 5
- 238000009360 aquaculture Methods 0.000 abstract description 3
- 244000144974 aquaculture Species 0.000 abstract 1
- 230000001580 bacterial effect Effects 0.000 description 15
- 229910002651 NO3 Inorganic materials 0.000 description 13
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 13
- 238000009395 breeding Methods 0.000 description 9
- 230000001488 breeding effect Effects 0.000 description 9
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 8
- 239000002609 medium Substances 0.000 description 8
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Inorganic materials [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 description 7
- 108020004465 16S ribosomal RNA Proteins 0.000 description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 230000035508 accumulation Effects 0.000 description 6
- 125000001477 organic nitrogen group Chemical group 0.000 description 6
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Substances [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 5
- 241000752227 Zobellella Species 0.000 description 5
- 238000012258 culturing Methods 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 241000251468 Actinopterygii Species 0.000 description 4
- 241000143060 Americamysis bahia Species 0.000 description 4
- 239000007836 KH2PO4 Substances 0.000 description 4
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 4
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 4
- 229910052564 epsomite Inorganic materials 0.000 description 4
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 4
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 4
- 229940074404 sodium succinate Drugs 0.000 description 4
- ZDQYSKICYIVCPN-UHFFFAOYSA-L sodium succinate (anhydrous) Chemical compound [Na+].[Na+].[O-]C(=O)CCC([O-])=O ZDQYSKICYIVCPN-UHFFFAOYSA-L 0.000 description 4
- 238000002798 spectrophotometry method Methods 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- 238000004659 sterilization and disinfection Methods 0.000 description 4
- 241000193830 Bacillus <bacterium> Species 0.000 description 3
- 239000002131 composite material Substances 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000007789 gas Substances 0.000 description 3
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 3
- 238000009630 liquid culture Methods 0.000 description 3
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 3
- 238000004321 preservation Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 235000013619 trace mineral Nutrition 0.000 description 3
- 239000011573 trace mineral Substances 0.000 description 3
- 241000588986 Alcaligenes Species 0.000 description 2
- UFWIBTONFRDIAS-UHFFFAOYSA-N Naphthalene Chemical compound C1=CC=CC2=CC=CC=C21 UFWIBTONFRDIAS-UHFFFAOYSA-N 0.000 description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 238000005374 membrane filtration Methods 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical compound OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 description 2
- 239000002351 wastewater Substances 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 241000588807 Bordetella Species 0.000 description 1
- 102000012286 Chitinases Human genes 0.000 description 1
- 108010022172 Chitinases Proteins 0.000 description 1
- 229910021580 Cobalt(II) chloride Inorganic materials 0.000 description 1
- 241001478240 Coccus Species 0.000 description 1
- 229910021592 Copper(II) chloride Inorganic materials 0.000 description 1
- RFSUNEUAIZKAJO-VRPWFDPXSA-N D-Fructose Natural products OC[C@H]1OC(O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-VRPWFDPXSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 102000001554 Hemoglobins Human genes 0.000 description 1
- 108010054147 Hemoglobins Proteins 0.000 description 1
- 208000032843 Hemorrhage Diseases 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Chemical compound OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 229910021380 Manganese Chloride Inorganic materials 0.000 description 1
- GLFNIEUTAYBVOC-UHFFFAOYSA-L Manganese chloride Chemical compound Cl[Mn]Cl GLFNIEUTAYBVOC-UHFFFAOYSA-L 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 229910018890 NaMoO4 Inorganic materials 0.000 description 1
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- 241001057811 Paracoccus <mealybug> Species 0.000 description 1
- 208000005374 Poisoning Diseases 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 241000926565 Zunongwangia endophytica Species 0.000 description 1
- CVTZKFWZDBJAHE-UHFFFAOYSA-N [N].N Chemical compound [N].N CVTZKFWZDBJAHE-UHFFFAOYSA-N 0.000 description 1
- 231100000570 acute poisoning Toxicity 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 239000005422 algal bloom Substances 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000010170 biological method Methods 0.000 description 1
- 210000000476 body water Anatomy 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- ORTQZVOHEJQUHG-UHFFFAOYSA-L copper(II) chloride Chemical compound Cl[Cu]Cl ORTQZVOHEJQUHG-UHFFFAOYSA-L 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 229960000935 dehydrated alcohol Drugs 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- DNJIEGIFACGWOD-UHFFFAOYSA-N ethanethiol Chemical compound CCS DNJIEGIFACGWOD-UHFFFAOYSA-N 0.000 description 1
- 238000012851 eutrophication Methods 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 230000001815 facial effect Effects 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 108060003552 hemocyanin Proteins 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 239000011565 manganese chloride Substances 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 229910052603 melanterite Inorganic materials 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000012543 microbiological analysis Methods 0.000 description 1
- LGQLOGILCSXPEA-UHFFFAOYSA-L nickel sulfate Chemical compound [Ni+2].[O-]S([O-])(=O)=O LGQLOGILCSXPEA-UHFFFAOYSA-L 0.000 description 1
- 229910000363 nickel(II) sulfate Inorganic materials 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- YBVAXJOZZAJCLA-UHFFFAOYSA-N nitric acid nitrous acid Chemical compound ON=O.O[N+]([O-])=O YBVAXJOZZAJCLA-UHFFFAOYSA-N 0.000 description 1
- 229910000069 nitrogen hydride Inorganic materials 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 231100000572 poisoning Toxicity 0.000 description 1
- 230000000607 poisoning effect Effects 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 239000004323 potassium nitrate Substances 0.000 description 1
- 235000010333 potassium nitrate Nutrition 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 229960004889 salicylic acid Drugs 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- WXMKPNITSTVMEF-UHFFFAOYSA-M sodium benzoate Chemical compound [Na+].[O-]C(=O)C1=CC=CC=C1 WXMKPNITSTVMEF-UHFFFAOYSA-M 0.000 description 1
- 239000004299 sodium benzoate Substances 0.000 description 1
- 235000010234 sodium benzoate Nutrition 0.000 description 1
- 235000010288 sodium nitrite Nutrition 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 1
- 229910000368 zinc sulfate Inorganic materials 0.000 description 1
- 239000011686 zinc sulphate Substances 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W10/00—Technologies for wastewater treatment
- Y02W10/10—Biological treatment of water, waste water, or sewage
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses Zhuo Beier Salmonella and its applications that one plant has nitrification and denitrification function simultaneously.Zhuo Beier Salmonella (Zobellella sp.) 2G-5, it is preserved in Guangdong Province's Culture Collection, address is No. 59 building of the compound of Xianlie Middle Rd., Yuexiu Zone, Guangzhou, Guangdong 100, the deposit date is on 03 06th, 2019, deposit number GDMCCNo.60606.Zhuo Beier Salmonella 2G-5 of the invention can quickly and efficiently remove ammonia nitrogen, nitrite nitrogen and nitrate nitrogen and its total nitrogen in water body, to above-mentioned three kinds inorganic nitrogen-sourced and total nitrogen removal effects 98% or more, and in the process without other inorganic nitrogen-sourced obvious accumulation, it is without secondary pollution, the bacterium has very big application value in cultivation water improvement, such as reduces the harm of ammonia nitrogen, nitrite nitrogen and nitrate nitrogen to cultivation object in aquaculture process.
Description
Technical field
The invention belongs to microbe applications and field of environment engineering technology, and in particular to one plant have simultaneously heterotrophic nitrification and
The Zhuo Beiershi denitrifying bacterium of aerobic denitrification function and its application in cultivation water improvement.
Background technique
It is commonly used with aquatic products intensive culture mode, in breeding process feed a part in cultivated animals digestion, inhale
Ammonia excretion is converted into breeding environment in the metabolic processes such as receipts, is also further converted into nitrite and nitrate, thus
Cause nitrogen content in cultivation water excessively high.Water element, which crosses high energy, leads to water eutrophication, and algal bloom consumes water
Dissolved oxygen in body causes the various aquatic animal mortalities such as fish so that water becomes stench.Such as excess of ammonia ammonia and nitrite
It is the main contributor for causing the aquiculture animals such as fishes and shrimps pathogenic.Studies have shown that excessive nitrite is to fish in breeding water body
Shrimp, which mainly passes through, influences its respiration, reduces the hemocyanin in the hemoglobin and shrimps blood in Fish Blood to oxygen
Affinity, to generate murder by poisoning to aquiculture animal.The excessive ammonia nitrogen of breeding water body (mainly has NH4 +With non-ionic NH3Two
Kind of form) immunocompetence of fishes and shrimps can be made to be forced to, the skin, stomach and enteron aisle facial mask of cultivated animals can be also destroyed, body surface and interior is caused
Portion's organ hemorrhage, so as to cause acute poisoning death.Therefore, it is dirty how cost-effectively to remove excessive nitrogen in breeding water body
Object is contaminated, a kind of efficient cultivation water modifying agent is found and has been to be concerned by more and more people.
Currently, mainly have chemistry, physics and biological method to the processing technique of nitrogen excessive in water body, wherein biological denitrificaion
Processing technique overcomes some defects of physical chemistry denitrogenation, has easy to operate, wide adaptation range, high treating effect, base
The advantages that without secondary pollution in sheet, the technology has been widely used in recent years.Microbial denitrogenation be studied in biological denitrificaion it is more
Method, conventionally, denitrification process is realized under anaerobic, and in recent years, is goed deep into to denitrifier
Research finds that some bacterium can have a heterotrophic nitrification and denitrifying function simultaneously under aerobic condition, and such bacterium is removing water body
There is unique advantage in terms of middle ammonia nitrogen, nitrite nitrogen and nitrate nitrogen, ammonia nitrogen or nitrate nitrogen can be finally reduced to gas or
Itself assimilates and is detached from breeding water body, noresidue, does not cause secondary pollution, improves and maintains breeding ecological balance, reduce disease
Occur, plays a significant role in cultivation water improvement.
There are diversity on Physiology and biochemistry and systematic growth for aerobic denitrifying bacteria.Existing research shows aerobic denitrification
Bacterium is distributed mainly on pseudomonad (Pseudomonas), Bacillus alcaligenes (Alcaligenes), secondary coccus (Paracoccus)
And in the category such as bacillus (Bacillus), and pertinent literature and patent are also more.Related Zhuo Beier Bordetella
(Zobellella) with the bacterial strain of denitrification in, the country's invention authorized at present only 2, patent 201510316321.7
(applying date 2015.06.10) discloses one plant of ocean Zhuo Beier Salmonella (Zobellella sp.) CYT1-8, with mode bacterium
The similar highest of Zobellella denitrificansZD1,16S rRNA similitude are 99.04%, bacterium involved by the patent
Strain can quickly remove NO3-N and NO2-N in water body under aerobic condition, but it has largely during removing nitrate
Nitrite accumulation.Patent 201310100699.4 (applying date 2013.08.07) discloses one plant of aerobic denitrifying bacteria Zhuo shellfish
Your Salmonella (Zobellella sp.) NKDN-01, and mode bacterium Zobellella taiwanensisZT1 similitude highest,
The bacterium has very high removal rate to high ammonia-nitrogen wastewater, it may have certain nitrate nitrogen removal effect, but it removes nitrite nitrogen
Except effect does not refer to, and in the patent, when handling artificial high ammonia-nitrogen wastewater to the bacterium, finally there is the increasing of micro nitrite nitrogen
Add.
Summary of the invention
The first purpose of the invention is to provide the Zhuo Beier that one plant has heterotrophic nitrification and aerobic denitrification function simultaneously
Salmonella (Zobellella sp.) 2G-5, is preserved in Guangdong Province's Culture Collection (GDMCC), address is Guangdong Province
No. 59 building of compound of XianLie Middle Road, Yuexiu District, GuangZhou City 100, the deposit date is on 03 06th, 2019, deposit number
GDMCCNo.60606。
The present invention, which separates, obtains one plant of Zhuo Beier Salmonella 2G-5 with heterotrophic nitrification and aerobic denitrification function, by bacterial strain
It is seeded to respectively with NH4 +- N, NO2 -- N and NO3 -- N is 28 DEG C in the culture medium of only nitrogen source, 180rpm shaken cultivation, respectively
When cultivating 0,12,18,24,36,48h, takes out culture solution and measures culture solution OD 600, while taking 2.0mL culture solution, be centrifuged,
Ammonia nitrogen, nitrite nitrogen and nitrate nitrogen in supernatant are measured, and measures total nitrogen in supernatant and thallus, as the result is shown Zhuo Beier Salmonella
2G-5 is respectively to the (NH in water body4)2SO4、NaNO2And KNO3All have removal effect rapidly and efficiently.It is being with nitrite nitrogen
In only nitrogen source culture medium, to NO after strain culturing 48h2 -The removal efficiency of-N and total nitrogen is 99.00% and 99.60%, wherein
27.08% inorganic nitrogen assimilates as thallus organic nitrogen；It is in only nitrogen source culture medium, after strain culturing 48h in nitrate nitrogen
To NO3 -The removal efficiency of-N and total nitrogen be 99.58% and 99.84%, wherein 29.72% inorganic nitrogen assimilate for thallus it is organic
Nitrogen；It is in only nitrogen source culture medium, to NH after strain culturing 48h in ammonia nitrogen4 +The removal efficiency of-N and total nitrogen is 98.72% He
99.80%, wherein 26.35% inorganic nitrogen assimilates as thallus organic nitrogen.Ammonia nitrogen, Asia in research bacterial strain 2G-5 removing water body
When nitrate nitrogen and nitrate nitrogen, the accumulation of apparent other forms inorganic nitrogen is not detected in the process.
Therefore, a second object of the present invention is to provide Zhuo Beier Salmonella 2G-5 in removal of ammonia and nitrogen, nitrite nitrogen and/or nitre
Application in state nitrogen.Such as applied in the preparation for preparing removal of ammonia and nitrogen, nitrite nitrogen and/or nitrate nitrogen.
It is preferred that being to carry out denitrogenation to the water body containing ammonia nitrogen, nitrite nitrogen and/or nitrate nitrogen, the application in water quality is improved.
The ammonia nitrogen is ammonium sulfate, and the nitrite nitrogen is sodium nitrite, and the nitrate nitrogen is potassium nitrate.
It is preferred that the described application is that Zhuo Beier Salmonella 2G-5 is seeded to nitrite nitrogen, nitrate nitrogen or ammonia nitrogen is unique
The application of growth and then denitrogenation in the culture medium of nitrogen source.
Third object of the present invention is to provide a kind of denitrogenation live bacteria agent with nitrification and denitrification function, (such as water quality changes
Good dose), contain Zhuo Beier Salmonella 2G-5 as active constituent.
Patent 201510316321.7 (applying date 2015.06.10) discloses one plant of ocean Zhuo Beier Salmonella
(Zobellella sp.) CYT1-8, to the similar highest of mode bacterium Zobellella denitrificansZD1,16S
RRNA similitude is 99.04%, and bacterial strain involved by the patent can quickly remove nitrate and nitrous in water body under aerobic condition
Hydrochlorate, but it has a large amount of nitrite accumulations during removing nitrate, and bacterial strain according to the present invention is to nitric acid
In salt removal process, thus the accumulation without obvious nitrite speculates bacterial strain of the present invention in denitrification process, nitrite is also
Original enzyme activity is higher, can soon by by nitrate reduction at nitrite be reduced further into nitrogenous gas, in reality
There is biggish application potential on breeding water body water quality improvement.
Zhuo Beier Salmonella 2G-5 of the invention can quickly and efficiently remove ammonia nitrogen, nitrite nitrogen and nitrate nitrogen in water body
And its total nitrogen, to above-mentioned three kinds inorganic nitrogen-sourced and total nitrogen removal effects 98% or more, and in the process without other inorganic nitrogens
The obvious accumulation in source, without secondary pollution, which has very big application value in cultivation water improvement, such as reduces and supports in aquatic products
The harm to cultivation object such as ammonia nitrogen, nitrite nitrogen and nitrate nitrogen during growing.
Zobellella sp.2G-5 was preserved in Guangdong Province's Culture Collection on 03 06th, 2019
(GDMCC), preservation address: No. 59 building of compound of Xianlie Middle Rd., Yuexiu Zone, Guangzhou, Guangdong 100, deposit number GDMCCNo:
60606。
Detailed description of the invention
Fig. 1 is colonial morphology of the Zhuo Beier Salmonella 2G-5 on solid plate；
Fig. 2 is the photo of Zhuo Beier Salmonella 2G-5 (100 ×) under an optical microscope；
Fig. 3 is Zhuo Beier Salmonella 2G-5 growth tendency and removing NO2 -The ability schematic diagram of-N；
Fig. 4 is Zhuo Beier Salmonella 2G-5 growth tendency and removing NO3 -The ability schematic diagram of-N；
Fig. 5 is Zhuo Beier Salmonella 2G-5 growth tendency and removing NH4 +The ability schematic diagram of-N.
Specific embodiment
The following examples are further illustrations of the invention, rather than limiting the invention.
Embodiment 1:
1, the enrichment of Zhuo Beier Salmonella (Zobellella sp.) 2G-5, isolate and purify and preservation
Enrichment culture:
Enriched medium is prepared, is formulated as follows: sodium succinate 2.5g, trisodium citrate dihydrate 2.5g, (NH4)2SO40.66g,KNO31.01g,NaNO20.69g, K2HPO41g, KH2PO41g, MgSO4·7H2O 0.2g, pH 7.4, is settled to
1000mL dispenses every bottle of 100mL into 250mL triangular flask.121 DEG C, 20min, autoclave sterilization.Then sterile mistake is added
Compounded carbons 1% (mass fraction) after filter, composite trace element 0.2% (mass fraction).Compounded carbons: D-Glucose
6.9g, D-Fructose 6.9g, D- lactose 6.9g, sodium acetate 9.5g, mass fraction 90% lactic acid 6.4mL, mannitol 7g, dehydrated alcohol
7mL, glycerol 6.3mL, sodium benzoate 4.8g, salicylic acid 4.6g are dissolved in 500mL water, and 7.4,0.22 μm of membrane filtration of pH removes
Bacterium.Micro-mixed liquor: ZnSO4·7H2O 0.1g, MnCl2·4H2O 0.4g, H3BO31.24g CoCl2·2H2O
0.5g, CuCl2·2H2O 0.5g, NiSO4·6H2O 0.02g, NaMoO4·2H2O 0.4g, KI 0.2g, CaCl25g,
FeSO4·7H2O 1.1g, EDTA-Na 10g, is dissolved in 1000mLH27.4,0.22 μm of membrane filtration of O, pH.
Shantou Nan ' ao Island Fish-shrimp mix-culture pond water sample 10mL is taken, access is equipped in the conical flask of above-mentioned enriched medium,
28 DEG C, 180rpm, 5d is cultivated, the enrichment of denitrifying bacteria is carried out.To the culture after above be incubated for, switching 1mL to new is equipped with
In the conical flask of enriched medium, 28 DEG C, 180rpm, 5d is cultivated, secondary enrichment is carried out, ibid carried out third time again and be enriched with training
It supports.
It isolates and purifies and preservation
For above-mentioned enrichment culture object three times, using method of dilution butteron on plate applying solid plate (solid medium used be
The agar of 1.7% (mass fraction) is added on the basis of enrichment culture based formulas)；Colonial morphology is visually observed, and purifying of crossing,
The purified bacterium colony of picking to 5mL pipe containing Du Shi fluid nutrient medium (enriched medium, but by three kinds it is inorganic nitrogen-sourced separate), 28
DEG C be incubated for observation produce gas situation, discovery bacterial strain 2G-5 can by Nitrite transformation be gaseous product.To bacterial strain after purification, expand
It cultivates and conservation glycerol tube and freeze-drying is managed.
Morphological feature
The morphological feature of the bacterial strain 2G-5 are as follows: Gram-negative bacteria；Colony colour is milky, translucent (see figure
1)；Bacterium colony is round, regular edges, and surface wettability is smooth；Cell shape is rod-short (see Fig. 2).The bacterial strain 2G-5 can divide
Not with (NH4)2SO4、NaNO2And KNO3To be grown in the culture medium of only nitrogen source.
The 16S rDNA of 2 Zhuo Bell Salmonella (Zobellella sp.) 2G-5 is identified
The genomic DNA for extracting bacterial strain 2G-5, is expanded with bacterial 16 S rRNA gene magnification universal primer 27F/1492R
To PCR product, and send to Science and Technology Ltd., Shanghai Major Biological Medical Technology Co., Ltd. (Guangzhou Branch) and carry out sequence, sequencing result
16S rDNA sequence in (nucleotide sequence of 16S rRNA is as shown in SEQ ID NO.1) and EzBioCloud site databases
Sequence analysis analysis is carried out, interpretation of result shows the 16S of itself and mode bacterium Zobellella denitrificansZD1
RRNA gene order similitude highest is 98.43%, with two plants of mode bacterium Z.taiwanensisZT1 and Z.aerophile
The similitude of JC2671 is 97.36%, and the similitude with mode bacterium Z.endophytica 59N8 is 97.21%.Based on
Upper interpretation of result, Preliminary Identification bacterial strain of the present invention are Zobellellasp., are named as Zhuo Beier Salmonella (Zobellellasp.)
2G-5, Guangdong Province's Culture Collection was preserved on 03 06th, 2019, and address is Guangzhou, Guangdong Yuexiu District
No. 59 building of compound of martyr Road 100, deposit number GDMCCNo.60606.
The nitrogen removal performance of 3 Zhuo Bell Salmonella (Zobellellasp.) 2G-5
It is inoculated with Zhuo Beier Salmonella 2G-5 [enrichment culture based formulas, but respectively with NO into fluid nutrient medium2 --N、NO3 --N
And NH4 +- N is only nitrogen source, specifically:
With NO2 -- N is the Liquid Culture based formulas of only nitrogen source: sodium succinate 2.5g, trisodium citrate dihydrate 2.5g,
NaNO20.69g, K2HPO41g, KH2PO41g, MgSO4·7H2O 0.2g, pH 7.4, is settled to 1000mL, and 121 DEG C, 20min,
Autoclave sterilization, the compounded carbons 1% (mass fraction, formula are same as above) after then addition is sterile filtered, composite trace element
0.2% (mass fraction, formula are same as above).
With NO3 -- N is the Liquid Culture based formulas of only nitrogen source: sodium succinate 2.5g, trisodium citrate dihydrate 2.5g,
KNO31.01g K2HPO41g, KH2PO41g, MgSO4·7H2O 0.2g, pH 7.4, is settled to 1000mL, and 121 DEG C, 20min,
Autoclave sterilization, the compounded carbons 1% (mass fraction, formula are same as above) after then addition is sterile filtered, composite trace element
0.2% (mass fraction, formula are same as above).
With NH4 +- N is the Liquid Culture based formulas of only nitrogen source: sodium succinate 2.5g, trisodium citrate dihydrate 2.5g,
(NH4)2SO40.66g, K2HPO41g, KH2PO41g, MgSO4·7H2O 0.2g, pH 7.4, is settled to 1000mL, and 121 DEG C,
20min, autoclave sterilization, the compounded carbons 1% (mass fraction, formula are same as above) after then addition is sterile filtered are compound micro-
Secondary element 0.2% (mass fraction, formula are same as above)],
28 DEG C, 180rpm shaking table culture, respectively culture 0,12,18,24,36, after 48h, sampling, a part of sample is used for
600 value of OD is detected, another part medium centrifugal takes supernatant, quantitative detection NO2 -、NO3 -、NH4 +With total nitrogen concentration (TN-N), this
Total nitrogen concentration (CN-N) in thallus is also measured outside.As a result such as Fig. 3, shown in 4,5, Zhuo Beier Salmonella 2G-5 is to NO in water body2 --N、
NO3 --N、NH4 +- N and total nitrogen therein have good removal effect.The bacterial strain is using nitrite nitrogen as the culture of only nitrogen source
In base, to NO after culture 48h2 -The removal efficiency of-N and total nitrogen is 99.00% and 99.60%, wherein 27.08% inorganic nitrogen is same
Turn to thallus organic nitrogen；In the culture medium that nitrate nitrogen is only nitrogen source, to NO after strain culturing 48h3 -- N and total nitrogen it is de-
Except rate is 99.58% and 99.84%, wherein 29.72% inorganic nitrogen assimilates as thallus organic nitrogen；It is only nitrogen source in ammonia nitrogen
In culture medium, to NH after strain culturing 48h4 +The removal efficiency of-N and total nitrogen is 98.72% and 99.80%, wherein 26.35%
Inorganic nitrogen assimilates as thallus organic nitrogen.And the bacterial strain ammonia nitrogen, nitrite nitrogen and when nitrate nitrogen in removing water body, in the process
The accumulation of apparent other forms inorganic nitrogen is not detected.Zhuo Beier Salmonella 2G-5 is one plant while having nitrification and denitrification function
The bacterium of energy, has very high removal effect to ammonia nitrogen, nitrite nitrogen and nitrate nitrogen in water body, and do not accumulate others in the process
Harmful nitrogen-containing products.In consideration of it, Zhuo Beier Salmonella 2G-5 is improved in cultivation water, the main nitrogen pollutant of breeding water body is removed,
Ensureing has huge potentiality in the practical application of the healthy growth of aquatic livestock.
Experimental determining method: NO2 -- N uses N- (1- naphthalene)-ethylendiamine dihydrochloride spectrophotometry；NO3 -- N is used
Determined by ultraviolet spectrophotometry nitrate；NH4 +- N uses Na Shi reagent spectrophotometry；Total nitrogen uses alkaline chitinase
Resolution-ultraviolet spectrophotometry.
Sequence table
<110>Guangdong Microbes Inst (microbiological analysis inspection center, Guangdong Province)
<120>one plants have Zhuo Beier Salmonella and its application of nitrification and denitrification function simultaneously
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1419
<212> DNA
<213>Zhuo Beier Salmonella (Zobellella sp.)
<400> 1
caacccactc ccatggtgtg acgggcggtg tgtacaaggc ccgggaacgt attcaccgca 60
acattctgat ttgcgattac tagcgattcc gacttcacgg agtcgagttg cagactccga 120
tccggactac gacgcgcttt ttgggattcg cttaccatcg ctggttcgcc gccctctgta 180
cgcgccattg tagcacgtgt gtagccctac ccgtaagggc catgatgact tgacgtcgtc 240
cccaccttcc tccggtttat caccggcagt ctcctttgag ttcccgaccg aatcgctggc 300
aacaaaggac aagggttgcg ctcgttgcgg gacttaaccc aacatctcac gacacgagct 360
gacgacagcc atgcagcacc tgtatgtaag ttcccgaagg cacccatcca tctctggaaa 420
gttcttacta tgtcaagggt aggtaaggtt cttcgcgttg catcgaatta aaccacatgc 480
tccaccgctt gtgcgggccc ccgtcaattc atttgagttt taaccttgcg gccgtactcc 540
ccaggcggtc aacttaacgc gttagctccg gaacccgcgc tcaaatggca cagactccaa 600
gttgacatcg tttacagcgt ggactaccag ggtatctaat cctgtttgct ccccacgctt 660
tcgcacctga gcgtcagtct ttggccaggg ggccgccttc gccactggta ttccttccga 720
tctctacgca tttcaccgct acaccggaaa ttctaccccc ctctccaaga ctctagcctg 780
ccagttccaa atgcagttcc caggttgagc ccggggcttt cacatctggc ttaacagacc 840
gcctgcgtgc gctttacgcc cagtcattcc gattaacgct tgcaccctcc gtattaccgc 900
ggctgctggc acggagttag ccggtgcttc ttctgtgggt aacgtcacag taagtgggta 960
ttcgccactc acctttcctc cccactgaaa gtgctttaca acccgaaggc cttcttcaca 1020
cacgcggcat ggctgcatca gggtttcccc cattgtgcaa tattccccac tgctgcctcc 1080
cgtaggagtc tgggccgtgt ctcagtccca gtgtggctgg tcatcctctc agaccagcta 1140
gagatcgtcg ccttggtgag ccattacctc accaaccagc taatctcact tgggctcatc 1200
caatcgcgca aggcccgaag gtcccctgct ttcccccgta gggcgtatgc ggtattagcc 1260
gtcgtttcca acggttatcc ccctcgactg ggcagatacc caagccttac tcacccgtcc 1320
gccgctcgtc agcaaagtag caagctaccc tctgttaccg ctcgacttgc atgtgttagg 1380
cctgccgcca gcgttcaatc tgagccatga tcaaactct 1419
Claims (7)
1. Zhuo Bell Salmonella (Zobellella sp.) 2G-5, deposit number GDMCC No.60606.
2. application of the Zhuo Bell Salmonella 2G-5 described in claim 1 in removal of ammonia and nitrogen, nitrite nitrogen and/or nitrate nitrogen.
3. application according to claim 2, which is characterized in that Zhuo Beier Salmonella 2G-5 is preparing removal of ammonia and nitrogen, nitrous state
It is applied in the preparation of nitrogen and/or nitrate nitrogen.
4. application according to claim 2, which is characterized in that Zhuo Beier Salmonella 2G-5 to containing ammonia nitrogen, nitrite nitrogen and/or
The water body of nitrate nitrogen carries out denitrogenation, improves the application in water quality.
5. application according to claim 2, which is characterized in that the described application be Zhuo Beier Salmonella 2G-5 is seeded to
Nitrite nitrogen, nitrate nitrogen or ammonia nitrogen are the application of growth and then denitrogenation in the culture medium of only nitrogen source.
6. a kind of denitrogenation live bacteria agent with nitrification and denitrification function, which is characterized in that contain tall and erect shellfish described in claim 1
That Salmonella 2G-5 is as active constituent.
7. denitrogenation live bacteria agent according to claim 6, which is characterized in that the denitrogenation live bacteria agent is improver of water quality.
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| CN113800652B (en) * | 2021-10-28 | 2022-07-26 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Salt-tolerant aerobic denitrifying bacterium and application of coupling activated carbon thereof in strengthening water body pollution treatment |
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