CN117887622A - Pseudomonas fragi without sugar decomposition and application thereof - Google Patents
Pseudomonas fragi without sugar decomposition and application thereof Download PDFInfo
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- CN117887622A CN117887622A CN202410004699.2A CN202410004699A CN117887622A CN 117887622 A CN117887622 A CN 117887622A CN 202410004699 A CN202410004699 A CN 202410004699A CN 117887622 A CN117887622 A CN 117887622A
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- 241000589538 Pseudomonas fragi Species 0.000 title description 2
- 238000000354 decomposition reaction Methods 0.000 title description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 29
- 239000002351 wastewater Substances 0.000 claims abstract description 22
- 230000000813 microbial effect Effects 0.000 claims abstract description 18
- 241000589771 Ralstonia solanacearum Species 0.000 claims abstract description 16
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 16
- MBMLMWLHJBBADN-UHFFFAOYSA-N Ferrous sulfide Chemical compound [Fe]=S MBMLMWLHJBBADN-UHFFFAOYSA-N 0.000 claims abstract description 11
- 238000009629 microbiological culture Methods 0.000 claims abstract description 3
- 238000000855 fermentation Methods 0.000 claims description 34
- 230000004151 fermentation Effects 0.000 claims description 34
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 24
- 238000000034 method Methods 0.000 claims description 19
- 239000000843 powder Substances 0.000 claims description 18
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 16
- 239000007788 liquid Substances 0.000 claims description 15
- 241000589516 Pseudomonas Species 0.000 claims description 14
- 238000012258 culturing Methods 0.000 claims description 14
- 239000001888 Peptone Substances 0.000 claims description 13
- 108010080698 Peptones Proteins 0.000 claims description 13
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 13
- 230000004913 activation Effects 0.000 claims description 13
- 235000019319 peptone Nutrition 0.000 claims description 13
- 238000011218 seed culture Methods 0.000 claims description 13
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 12
- 229910052799 carbon Inorganic materials 0.000 claims description 12
- 229910052757 nitrogen Inorganic materials 0.000 claims description 12
- 241000222122 Candida albicans Species 0.000 claims description 8
- 229940095731 candida albicans Drugs 0.000 claims description 8
- 235000015097 nutrients Nutrition 0.000 claims description 8
- 239000011780 sodium chloride Substances 0.000 claims description 8
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 7
- 239000008103 glucose Substances 0.000 claims description 7
- 150000004763 sulfides Chemical class 0.000 claims description 7
- 230000007613 environmental effect Effects 0.000 claims description 6
- 238000011081 inoculation Methods 0.000 claims description 6
- 239000002054 inoculum Substances 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 4
- 241001078784 Pseudochrobactrum asaccharolyticum Species 0.000 claims description 4
- 238000005273 aeration Methods 0.000 claims description 4
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 claims description 4
- 239000002904 solvent Substances 0.000 claims description 4
- 241000235646 Cyberlindnera jadinii Species 0.000 claims description 3
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 3
- 229920002472 Starch Polymers 0.000 claims description 3
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 3
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 3
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 3
- 230000000593 degrading effect Effects 0.000 claims description 3
- 239000001301 oxygen Substances 0.000 claims description 3
- 229910052760 oxygen Inorganic materials 0.000 claims description 3
- 239000001632 sodium acetate Substances 0.000 claims description 3
- 235000017281 sodium acetate Nutrition 0.000 claims description 3
- 239000008107 starch Substances 0.000 claims description 3
- 235000019698 starch Nutrition 0.000 claims description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 2
- 229930006000 Sucrose Natural products 0.000 claims description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 2
- 229940041514 candida albicans extract Drugs 0.000 claims description 2
- 239000004202 carbamide Substances 0.000 claims description 2
- 235000013877 carbamide Nutrition 0.000 claims description 2
- 239000004323 potassium nitrate Substances 0.000 claims description 2
- 235000010333 potassium nitrate Nutrition 0.000 claims description 2
- 229940074404 sodium succinate Drugs 0.000 claims description 2
- ZDQYSKICYIVCPN-UHFFFAOYSA-L sodium succinate (anhydrous) Chemical compound [Na+].[Na+].[O-]C(=O)CCC([O-])=O ZDQYSKICYIVCPN-UHFFFAOYSA-L 0.000 claims description 2
- 239000005720 sucrose Substances 0.000 claims description 2
- 239000012138 yeast extract Substances 0.000 claims description 2
- 239000004480 active ingredient Substances 0.000 claims 1
- 229920001542 oligosaccharide Polymers 0.000 claims 1
- 150000002482 oligosaccharides Chemical class 0.000 claims 1
- 238000005067 remediation Methods 0.000 claims 1
- 230000001523 saccharolytic effect Effects 0.000 claims 1
- UCKMPCXJQFINFW-UHFFFAOYSA-N Sulphide Chemical compound [S-2] UCKMPCXJQFINFW-UHFFFAOYSA-N 0.000 abstract description 17
- 239000013049 sediment Substances 0.000 abstract description 10
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 abstract description 8
- 230000015556 catabolic process Effects 0.000 abstract description 6
- 238000006731 degradation reaction Methods 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 5
- 230000006872 improvement Effects 0.000 abstract description 4
- 230000009286 beneficial effect Effects 0.000 abstract description 3
- 238000004321 preservation Methods 0.000 abstract description 3
- 239000002609 medium Substances 0.000 description 23
- 239000000243 solution Substances 0.000 description 23
- 230000001580 bacterial effect Effects 0.000 description 13
- 241000894006 Bacteria Species 0.000 description 12
- 238000002474 experimental method Methods 0.000 description 12
- 230000012010 growth Effects 0.000 description 11
- 239000001963 growth medium Substances 0.000 description 10
- 239000012153 distilled water Substances 0.000 description 7
- 229910017053 inorganic salt Inorganic materials 0.000 description 7
- 229910052717 sulfur Inorganic materials 0.000 description 7
- 239000011593 sulfur Substances 0.000 description 7
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 6
- 230000003213 activating effect Effects 0.000 description 6
- 230000008569 process Effects 0.000 description 6
- 239000010865 sewage Substances 0.000 description 6
- 239000010802 sludge Substances 0.000 description 6
- 230000001954 sterilising effect Effects 0.000 description 6
- 241000196324 Embryophyta Species 0.000 description 5
- 239000003344 environmental pollutant Substances 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
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- 238000002835 absorbance Methods 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 238000004659 sterilization and disinfection Methods 0.000 description 4
- 239000000758 substrate Substances 0.000 description 4
- 108020004465 16S ribosomal RNA Proteins 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 230000003698 anagen phase Effects 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 238000004108 freeze drying Methods 0.000 description 3
- 238000002955 isolation Methods 0.000 description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 3
- 235000019341 magnesium sulphate Nutrition 0.000 description 3
- 230000001590 oxidative effect Effects 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- -1 sulfur ion Chemical class 0.000 description 3
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 108010073771 Soybean Proteins Proteins 0.000 description 2
- 235000015278 beef Nutrition 0.000 description 2
- 239000011575 calcium Substances 0.000 description 2
- 239000001110 calcium chloride Substances 0.000 description 2
- 229910001628 calcium chloride Inorganic materials 0.000 description 2
- ZCCIPPOKBCJFDN-UHFFFAOYSA-N calcium nitrate Chemical compound [Ca+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O ZCCIPPOKBCJFDN-UHFFFAOYSA-N 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 239000003337 fertilizer Substances 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 239000011777 magnesium Substances 0.000 description 2
- YIXJRHPUWRPCBB-UHFFFAOYSA-N magnesium nitrate Chemical compound [Mg+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O YIXJRHPUWRPCBB-UHFFFAOYSA-N 0.000 description 2
- 239000011572 manganese Substances 0.000 description 2
- 229940099596 manganese sulfate Drugs 0.000 description 2
- 239000011702 manganese sulphate Substances 0.000 description 2
- 235000007079 manganese sulphate Nutrition 0.000 description 2
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 229960004249 sodium acetate Drugs 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 235000019710 soybean protein Nutrition 0.000 description 2
- 239000011573 trace mineral Substances 0.000 description 2
- 235000013619 trace mineral Nutrition 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 description 1
- 229920002774 Maltodextrin Polymers 0.000 description 1
- 239000005913 Maltodextrin Substances 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
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- 244000046052 Phaseolus vulgaris Species 0.000 description 1
- 241000588769 Proteus <enterobacteria> Species 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- CKUAXEQHGKSLHN-UHFFFAOYSA-N [C].[N] Chemical compound [C].[N] CKUAXEQHGKSLHN-UHFFFAOYSA-N 0.000 description 1
- 230000001133 acceleration Effects 0.000 description 1
- 238000003916 acid precipitation Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 1
- 235000011114 ammonium hydroxide Nutrition 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 238000005352 clarification Methods 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 229960001031 glucose Drugs 0.000 description 1
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 229910000037 hydrogen sulfide Inorganic materials 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 239000012263 liquid product Substances 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 229940035034 maltodextrin Drugs 0.000 description 1
- 239000011565 manganese chloride Substances 0.000 description 1
- 235000002867 manganese chloride Nutrition 0.000 description 1
- 229940099607 manganese chloride Drugs 0.000 description 1
- MIVBAHRSNUNMPP-UHFFFAOYSA-N manganese(2+);dinitrate Chemical compound [Mn+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O MIVBAHRSNUNMPP-UHFFFAOYSA-N 0.000 description 1
- ISPYRSDWRDQNSW-UHFFFAOYSA-L manganese(II) sulfate monohydrate Chemical compound O.[Mn+2].[O-]S([O-])(=O)=O ISPYRSDWRDQNSW-UHFFFAOYSA-L 0.000 description 1
- CDUFCUKTJFSWPL-UHFFFAOYSA-L manganese(II) sulfate tetrahydrate Chemical compound O.O.O.O.[Mn+2].[O-]S([O-])(=O)=O CDUFCUKTJFSWPL-UHFFFAOYSA-L 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 235000013379 molasses Nutrition 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 239000005416 organic matter Substances 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 238000000053 physical method Methods 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000005057 refrigeration Methods 0.000 description 1
- 239000012266 salt solution Substances 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000012807 shake-flask culturing Methods 0.000 description 1
- 229940080313 sodium starch Drugs 0.000 description 1
- 239000012265 solid product Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 229940032147 starch Drugs 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 229960004793 sucrose Drugs 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 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
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a pseudomonas solanacearum which does not decompose sugar and application thereof, and the strain is preserved in the China general microbiological culture Collection center with the address: the collection number of the national institute of microbiology, national academy of sciences, no. 3, north Chen West Lu 1, chao yang, beijing, is: CGMCC No.28059, the preservation date is: the invention provides the pseudomonas solanacearum for sugar-free, which has the beneficial effects that: the sulfide has strong degradation capability and quick effect, can efficiently and quickly degrade ferrous sulfide existing in the wastewater, oxidize the ferrous sulfide into white elemental sulfur, effectively solve the problem of black and stink of water and river sediment, and can be further used as a microbial agent for river sediment improvement.
Description
Technical Field
The invention relates to a pseudomonas solanacearum which does not decompose sugar, a microbial agent containing the pseudomonas solanacearum and application thereof in the field of environmental treatment, and belongs to the technical field of environmental microorganisms.
Background
Along with the acceleration of urban and industrialized processes, the sewage discharge amount is increasing, and the self-cleaning capability of urban water bodies is seriously affected by a large amount of sewage discharge.
The disposal of these high concentration and difficult to degrade sulfur-containing organic wastewater becomes a new challenge for these industries where more and more sulfide generated during the production process is dissolved in the wastewater for disposal. The concentration of the sulfur-containing compounds in the wastewater is high, the toxicity is high, the environmental pollution is serious, natural disasters are further caused, and even the human health is endangered, such as acid rain.
The emission of pollutants ensures that the organic matter content in river water is high, and the total nitrogen of ammonia nitrogen is greatly out of standard, so that the water quality of a river channel is rapidly deteriorated, even the river channel is smelly and blackened. River dredging is a main measure for guaranteeing water resource safety and good water environment at present, but how to treat river sediment after dredging is still a social problem. Because the sediment contains a large amount of organic matters and nutrient substances, the sediment is stacked or improperly treated at will, and secondary pollution is easily caused to the water body. The pollutant in the bottom mud exceeds the standard and cannot be reused as farmland fertilizer, so the importance of bottom mud improvement gradually begins to attract the attention of environmental protection people.
The main reason for the black and odorous substrate sludge is that after the corpse and the excrement of the aquatic organisms are deposited, harmful bacteria are utilized in the anaerobic environment of the substrate sludge to generate hydrogen sulfide gas, and the gas overflows to generate odor and simultaneously black ferrous sulfide sediment is generated in the water body.
There are many common ways of substrate sludge modification, such as physical modification, chemical modification and microbiological modification. Common physical improvement methods include screening, compression and the like, and impurities and pollutants in the bottom mud can be removed through treatment of the physical method, so that the aim of purifying the water body is fulfilled. The chemical method is to adjust the acid-base balance by adding an oxidant and a reducing agent, so as to promote the chemical reaction of organic matters and pollutants in the sediment to be removed. However, these two methods are generally faced with the problems of high cost, large amount of manpower and material resources are required for pushing, and secondary pollution to the environment may be caused by the use and residue of chemical reagents. Therefore, the application of the microbial treatment method is becoming more and more widespread.
The microbial agent is combined with beneficial bacteria carried by the river channel, so that dominant microbial bacteria can be quickly established in the original environment of the water body, various organic matters and pollutants are converted, decomposed and discharged, the water body is purified, the destroyed ecological system in the water body system is restored, and the original ecological function of the bottom mud is recovered. The treated bottom mud can be used as fertilizer for watering, thereby realizing secondary utilization.
In conclusion, through separating sulfur oxidizing bacteria with stronger sulfur ion oxidation function, free sulfur ions can be converted into sulfur simple substances which are insoluble in water and can be settled by utilizing the metabolism mechanism of microorganisms under anaerobic, anoxic and aerobic conditions, so that the purposes of improving substrate sludge and improving black and odorous water body are achieved.
Disclosure of Invention
Aiming at the defects of the existing treatment method of sulfide in wastewater and the existing sediment improvement method, the invention provides a pseudomonas solanacearum which does not decompose sugar, a microbial agent containing the pseudomonas solanacearum and application thereof, and the pseudomonas solanacearum can degrade sulfide in wastewater and ferrous sulfide sediment which causes the sediment to turn black by utilizing a self metabolic mechanism.
The strain of the pseudomonas solanacearum (Pseudochrobactrum asaccharolyticum) BC-1 is preserved in the China general microbiological culture Collection center with the address of: the collection number of the national institute of microbiology, national academy of sciences, no. 3, north Chen West Lu 1, chao yang, beijing, is: CGMCC No.28059, the preservation date is: the 16S rDNA sequence of the strain is shown as SEQ ID No. 1 in 28 days 7 of 2023, and the non-sugar-decomposing pseudomonas palustris referred to as the non-sugar-decomposing pseudomonas palustris (Pseudochrobactrum asaccharolyticum) BC-1 strain.
The invention provides the beneficial effects of the non-sugar-decomposing pseudomonas palustris that:
(1) The sulfide has strong degradation capability and quick effect, and the maximum 24h degradation rate can reach 96 percent for the initial sulfide concentration of about 100mg/L under the condition of 30 ℃.
(2) Ferrous sulfide existing in the wastewater can be efficiently and rapidly degraded, and oxidized into white elemental sulfur, so that the black and odorous problems of water bodies and river bottom mud are effectively solved, and the ferrous sulfide can be used as a microbial agent for improving the river bottom mud;
(3) The strain has the advantages of simple culture method, strong environmental adaptability, rapid propagation and high safety.
The invention also claims a microbial agent containing the pseudomonas solanacearum.
The invention also claims a fermentation method of the non-sugar-decomposing pseudomonas, which comprises the following steps:
(1) Primary seed culture: inoculating the non-sugar-decomposing pseudomonas into a nutrient medium under the aseptic condition, and culturing for 12-24 hours at 25-35 ℃ and 150-300rpm to obtain a first-stage seed culture solution;
(2) Secondary seed culture: inoculating the primary seed culture solution into a nutrient medium according to an inoculum size of 1-10vol% under a sterile condition, and culturing for 12-24h at 25-35 ℃ and 150-300rpm to obtain a secondary seed culture solution;
(3) Fermentation: after the fermentation medium is sterilized, inoculating the secondary seed culture solution into the fermentation medium according to the inoculum size of 1-10vol%, controlling the temperature to be 25-35 ℃, the rotating speed to be 150-300rpm, fermenting under the condition that the aeration ratio is 1 (1-2), and stopping fermenting when dissolved oxygen starts to rise to obtain the fermentation liquid.
The aeration ratio in the fermentation process of the present invention refers to the ratio of the volume of air introduced into the fermenter per minute to the total volume of fermentation broth.
The nutrient medium comprises the following components: 5-15g/L peptone, 3-8g/L yeast powder, 8-12g/L sodium chloride, water as solvent, and pH=6.5-7.5.
The fermentation medium comprises the following components: 5-15g/L of carbon source and 3-8g/L, PO of nitrogen source 4 3 -1-3g/L、K + 1-2g/L、Mg 2+ 0.05-0.10g/L、Ca 2+ 0.5-1.5g/L、Mn 2+ 0.05-0.1g/L, water as solvent, pH=6-8.
Preferably, the PO 4 3 -and K + The source of (a) is dipotassium hydrogen phosphate or monopotassium phosphate, and the Mg 2+ The source of the Ca is one or more of magnesium sulfate, magnesium nitrate or magnesium chloride 2+ Is one or more of calcium chloride and calcium nitrate, and Mn 2+ The source of the catalyst is one or more of manganese sulfate monohydrate, manganese sulfate tetrahydrate, manganese nitrate or manganese chloride.
Further, the carbon source is selected from one or more of glucose, sucrose, starch, sodium acetate or sodium succinate.
Further, the nitrogen source is selected from one or more of yeast extract, peptone, urea, ammonium sulfate or potassium nitrate.
In the practical application process, the form of the final product of the non-sugar-decomposing pseudomonas can be determined according to the practical use and storage requirements, when the liquid product is required to be used, the fermentation liquor is diluted to the required concentration to be directly used, when the solid product is required to be used, the fermentation liquor can be centrifuged to obtain bacterial mud, and then the solid bacterial powder is prepared by adopting a freeze-drying process.
The invention also claims a method for purifying sewage and wastewater by using the activation solution of the non-sugar-decomposing candida albicans or the microbial agent containing the non-sugar-decomposing candida albicans, which comprises the step of inoculating the activation solution of the non-sugar-decomposing candida albicans or the microbial agent containing the non-sugar-decomposing candida albicans into the sewage and wastewater.
Further, the concentration of sulfide in the sewage water is 400mg/L or less, preferably 300mg/L or less, and most preferably 200mg/L or less.
Further, the inoculation amount of the activation solution of the non-sugar-decomposing Bacillus pseudochrous or the microbial agent containing the non-sugar-decomposing Bacillus pseudochrous is 100ppm or more, preferably 100 to 1000ppm.
Further, the temperature during the purification process is 25-35 ℃.
The invention also claims the application of the pseudomonas solanacearum and the microbial agent containing the pseudomonas solanacearum in the field of environmental treatment.
Preferably, the non-sugar-degrading pseudomonas solanacearum and the microbial agent containing the non-sugar-degrading pseudomonas solanacearum are used for degrading sulfides, more preferably, the sulfides are ferrous sulfide and H 2 S、HS - And S is 2- 。
Drawings
FIG. 1 is a graph showing the growth of the E.parahaemolyticus measured in example 2;
FIG. 2 is a plant wastewater containing a large amount of ferrous sulfide black precipitate;
FIG. 3 shows the factory wastewater after culturing for 72 hours by inoculating the non-sugar decomposing Pseudomonas BC-1.
Detailed Description
The principles and features of the present invention are described below in connection with examples, which are set forth only to illustrate the present invention and not to limit the scope of the invention.
Example 1 isolation and purification and identification of the sugar-free Pseudomonas
(1) Screening and isolation of strains:
collecting sludge and sludge suspension of a chemical plant, inoculating 10mL of sludge-water mixture into a 250mL headspace bottle containing 100mL of enrichment medium (yeast powder 5g, peptone 10g, sodium chloride 10g, distilled water 1L, pH=7.0, and sterilization at 121 ℃ for 20 min), and shaking-culturing on a constant temperature shaking table at 30 ℃ for 200r/min for one week to obtain enrichment liquid.
Taking the enriched liquid to dilute to 10 step by step -3 ，10 -4 ，10 -5 And 10 -6 After doubling, each dilution was applied to SOB solid selection medium (KH 2 PO 4 1g，NH 4 Cl 0.8g，MgCl 2 ·6H 2 O 0.8g，CaCl 2 ·2H 2 O0.01g，FeCl 3 ·6H 2 O 0.01g，MnCl 2 ·4H 2 O 0.04g，Na 2 S·9H 2 O1.2 g, beef extract 2g, peptone 10g, agar 20g, distilled water 1L, pH=7.0,sterilizing at 121deg.C for 20 min), culturing the coated plate in a constant temperature incubator at 30deg.C until single colony is grown, selecting single colony with different forms, transferring to test tube slant culture medium, culturing at 30deg.C for about 48 hr, and transferring to refrigerator at 4deg.C for preservation.
6 strains named BC-1, BC-2, BC-3, BC-4, BC-5 and BC-6 were obtained by co-isolation according to the above method.
(2) Evaluation of strains
The obtained 6 strains were inoculated into an activation medium (yeast powder 5g, peptone 10g, sodium chloride 10g, distilled water 1L, pH=7.0), cultured for 48 hours at 30℃in a shaker 200r/min to obtain an activation solution, and then 100mL of a liquid selection medium (KH 2 PO 4 1g，NH 4 Cl 0.8g，MgCl 2 ·6H 2 O 0.8g，CaCl 2 ·2H 2 O 0.01g，FeCl 3 ·6H 2 O 0.01g，MnCl 2 ·4H 2 O0.04g，Na 2 S·9H 2 0.4g of O, 2g of beef extract, 10g of peptone, 1L of distilled water, pH=7.0 and sterilization at 121 ℃ for 20 min) were inoculated into headspace bottles (250 mL), the respective strains were inoculated with an activating solution of 100ppm, and cultured at 200r/min on a shaker at 30 ℃, and the changes in sulfide content after 0, 24, 48, 72, 96 and 120 hours were measured using a LH-S3H sulfide meter manufactured by Beijing Lianhua Yongxing technology Co., ltd., in accordance with the special measuring reagent, and the results are shown in Table 1.
TABLE 1 degradation of sulfides by the strains obtained by the screening
| BC-1 | BC-2 | BC-3 | BC-4 | BC-5 | BC-6 | |
| 24h | 97.92 | 269.01 | 269.55 | 272.00 | 217.60 | 269.28 |
| 48h | 8.16 | 268.74 | 272.00 | 269.28 | 225.22 | 269.01 |
| 72h | 0.00 | 268.74 | 272.00 | 272.00 | 214.61 | 272.00 |
| 96h | 0.00 | 267.65 | 271.18 | 263.02 | 209.44 | 268.46 |
| 120h | 0.00 | 268.46 | 272.00 | 265.74 | 207.81 | 270.10 |
As can be seen from the data in Table 1, the BC-1 strain has a fast effect and a good effect on removal of sulfides compared with other strains, the sulfide concentration of about 270ppm in the culture medium is evaluated, the complete removal can be achieved after 72 hours, the strain is re-inoculated into LB culture medium to prepare an activating solution, and then the activating solution is preserved at a temperature of-80 ℃.
(3) Detection and identification
The bacterial slant is detected and identified by a 16S rDNA gene sequence, and the identification result is that the bacterial slant is the pseudomonas solanacearum (Pseudochrobactrum asaccharolyticum). The sequence of the 16S rDNA gene of the strain is shown as SEQ ID No. 1.
EXAMPLE 2 exploration of growth conditions of the Pseudomonas solani without sugar
(1) Determination of optimal carbon source for Strain growth
The experimental method comprises the following steps:
preparing a fermentation medium, uniformly using yeast powder as a nitrogen source, respectively using glucose, molasses, brown sugar, maltodextrin, sodium acetate, starch and corn meal as carbon sources for experiments, and proportioning according to a carbon-nitrogen ratio of 2:1 and 1 g/L. Two inorganic salt solutions were prepared, the experiment was divided into two groups, and the concentration of each trace element in the fermentation medium using inorganic salt 1 was: dipotassium hydrogen phosphate 0.3%, magnesium sulfate 0.2%, manganese sulfate 0.2%, calcium carbonate 0.2%, and the concentration of each trace element in the fermentation medium using inorganic salt 2 is: 0.5% of magnesium sulfate, 0.3% of manganese sulfate, 1.5% of calcium chloride and 1.5% of dipotassium hydrogen phosphate. And increasing NA and LB culture medium experiment groups simultaneously, and searching an optimal culture medium of the non-sugar-dissolving pseudomonas palustris seed solution.
Respectively inoculating BC-1 strain, culturing at 30deg.C and 180r/min on shaking table, and periodically measuring OD 600 (OD 600 Represents the absorbance of the solution at 600nm, and the concentration of the bacterial culture is measured by using the absorbance, which is commonly used to refer to the cell density of the cells, and the growth of bacteria can be measured by OD 600 The values of (1) and the number of viable bacteria were used to determine the degree of growth of the strain, and the results are shown in Table 2, wherein the numbers 1 and 2 after the carbon source in Table 2 represent the ratio of inorganic salt 1 or inorganic salt 2 used in the formulation of inorganic salt in the medium.
TABLE 2 OD of the BC-1 Strain cultured in Medium of different carbon sources 600 Value and viable count
As is clear from the data in Table 2, the BC-1 strain grows to an optimal extent when the carbon source in the medium is glucose, and thus glucose is determined to be an optimal carbon source for the E.pseudochrous not saccharolyticus.
(2) Determination of optimal Nitrogen Source for Strain growth
The experimental method comprises the following steps:
preparing a fermentation medium, uniformly using glucose as a carbon source, and respectively using soybean protein isolate, soybean protein, yeast powder, peptone, ammonium sulfate and bean cake powder as nitrogen sources for experiments. Carbon to nitrogen ratio 2:1, inorganic salt was used in formulation 2.
Respectively inoculating BC-1 strain, culturing at 30deg.C and 180r/min on shaking table, and periodically measuring OD 600 (OD 600 Represents the absorbance of the solution at 600nm, and the concentration of the bacterial culture is measured by using the absorbance, which is commonly used to refer to the cell density of the cells, and the growth of bacteria can be measured by OD 600 Is monitored) and the number of viable bacteria to determine the degree of growth of the bacterial species, the results are shown in table 3.
TABLE 3 OD of the BC-1 Strain cultured in Medium with different Nitrogen sources 600 Value and viable count
As can be seen from the data in Table 3, the BC-1 strain grows to an optimal extent when the nitrogen source in the medium is yeast powder, and thus the yeast powder is determined to be the optimal nitrogen source of the Candida utilis without saccharolytica.
(3) Strain growth curve determination
The experimental method comprises the following steps:
and (3) inoculating the slant to an LB culture medium, performing shake culture for 48 hours to obtain an activating solution, and inoculating the activating solution to a new LB culture medium to start experiments, wherein the inoculation amount is 2%.
The experiment was divided into two experimental groups, each of which was made in three replicates. Nine am inoculation experiment group one, three in total. And (5) placing the first-stage activating solution after inoculation into a refrigerator for refrigeration. OD (optical density) 600 The initial value is measured. OD of experiment group I 600 The measurement time is 9,11 am and 1,3,5 pm, the dilution factor is ten times. Five-point at night vaccinated experimental group two, three in total. OD (optical density) 600 Initial measurement, OD 600 The measurement time of (2) was 5,7 days later and 9,11,1,3,5 days next, and the dilution factor was ten times. After each sampling, the bacteria were stained or not by microscopic examination, and the results are shown in Table 4 and FIG. 1.
TABLE 4 growth Curve measurement data for BC-1 Strain
| Cultivation time (h) | OD 600 | Cultivation time (h) | OD 600 |
| 0 | 0.027 | 30 | 0.393 |
| 2 | 0.044 | 32 | 0.410 |
| 4 | 0.124 | 40 | 0.537 |
| 6 | 0.195 | 42 | 0.534 |
| 8 | 0.222 | 48 | 0.648 |
| 22 | 0.244 | 50 | 0.650 |
| 24 | 0.244 | 52 | 0.653 |
| 26 | 0.332 | 54 | 0.657 |
| 28 | 0.361 |
As can be seen from Table 4 and FIG. 1, the logarithmic growth phase of the non-saccharogenic pseudo-pallidum bacteria is about 30 to 40 hours. And (3) injection: the measurement of the growth curve may not be entirely accurate and the experimental logarithmic growth phase may be longer than the actual growth phase.
Example 3 production and post-treatment Process of Proteus parahaemolyticus
(1) Strain fermentation
1) First-stage shaking activation
In a sterile environment, the 1-cycle non-sugar decomposing pseudomonas BC-1 strain is selected and inoculated into a 250mL triangular flask filled with 100mL of enrichment medium (5 g of yeast powder, 10g of peptone, 10g of sodium chloride, pH=7.0 and sterilization at 121 ℃ for 20 min), and the mixture is placed at 30 ℃ for culturing for 24h under 180r/min to obtain a first-stage activation solution.
2) Two-stage shake flask culture
In a sterile environment, the primary activation solution is respectively transferred into 10mL to four 1L triangular flasks which are filled with 500mL of enrichment culture medium (yeast powder 5g, peptone 10g, sodium chloride 10g, distilled water 1L, pH=7.0 and sterilization at 121 ℃ for 20 min), and the culture is carried out for 24h at 30 ℃ and 180r/min to obtain the secondary activation solution.
3) 20L tank fermentation culture
Sterilizing the fermentation medium at 121 ℃ for 20min, wherein the formula of the fermentation medium is as follows: glucose 10g/L, yeast powder 5g/L, mgSO 4 0.4g/L，MnSO 4 0.2g/L，CaCl 2 3g/L，K 2 HPO 4 3g/L, inoculating the secondary seed culture solution into the fermentation medium according to an inoculum size of 5-10vol%, fermenting tank liquid amount of 70%, adjusting initial pH=7.0 with sodium hydroxide, and adjusting aeration ratio to 1:1.25 (m 3 ·min/m 3 ) And (3) fermenting and culturing at 180rpm and 30 ℃, wherein the pH=6.5 is controlled by adding ammonia water in the fermentation process, the fermentation period is about 40 hours, when dissolved oxygen starts to rise back, the fermentation is stopped immediately, the fermentation is at the end of the logarithmic phase, the viable count is up to 300 hundred million cfu/mL, the activity of the bacteria is strongest, the residual of fermentation nutrient substances is minimum, and the attenuation of the stored viable count is less.
2. Post-treatment process of fermentation liquor
In order to prepare the bacterial powder, the fermentation liquor can be treated by adopting a freeze-drying post-treatment process, and the viable count of the bacterial powder after freeze-drying can reach 2 trillion CFU/g.
EXAMPLE 4 evaluation of waste Water from Pseudomonas saccharolytica
Inoculating single bacterial colony of the non-saccharogenic candida albicans BC-1 into an activation culture medium (5 g of yeast powder, 10g of peptone, 10g of sodium chloride, 1L of distilled water and pH=7.0), and culturing for 48 hours at the temperature of 30 ℃ on a shaking table 200r/min to obtain an activation solution, wherein the active bacterial amount of the activation solution is 60 hundred million CFU/mL.
The sulfide oxidizing effect of the BC-1 strain was repeatedly evaluated using sulfur-containing wastewater from two plants, the initial sulfide concentration of the plant wastewater being 90mg/L and 119mg/L. Then, a headspace bottle (250 mL) containing 200mL of factory sewage was prepared, the activated liquid of the strain was inoculated into the headspace bottle, the inoculation amount of the activated liquid was 100ppm, the strain was cultured at a temperature of 30 ℃ in a shaker of 200r/min, and the change of the sulfide content after 0, 24, 48 and 72 hours was measured by using an LH-S3H sulfide meter produced by Beijing Lianhua Yongxing technology development Co., ltd in combination with a special measuring reagent, and the results are shown in Table 5.
TABLE 5 degradation of sulfides in plant wastewater by BC-1 strains
EXAMPLE 5 evaluation of ferrous sulfide degradation of Pseudomonas saccharolytica
Inoculating single bacterial colony of the non-saccharogenic candida albicans BC-1 into an activated liquid culture medium (5 g of yeast powder, 10g of peptone, 10g of sodium chloride, 1L of distilled water and pH=7.0), and culturing for 48 hours at the temperature of 30 ℃ on a shaking table 200r/min to obtain an activated liquid, wherein the live bacterial amount of the activated liquid is 60 hundred million CFU/mL.
Then, a headspace bottle (250 mL) containing 250mL of factory wastewater was prepared, as shown in FIG. 2, the activated liquid of the strain was inoculated into the headspace bottle, the inoculum size of the activated liquid was 100ppm, and the culture was carried out under the conditions of 200r/min and 30℃on a shaker, and the change of the liquid in the bottle was observed. The content of sulfide after clarification of water is measured by using an LH-S3H sulfide measuring instrument produced by Beijing Lianhua Yongxing technology development Co., ltd, and the result after cultivation for 72 hours is shown in FIG. 3.
Experiments prove that after the activated liquid inoculated with the BC-1 strain in the factory wastewater is cultured, ferrous sulfide in the wastewater is oxidized into white elemental sulfur by the functional bacteria, and the concentration of sulfide in the wastewater is measured to be zero.
The foregoing description of the preferred embodiments of the invention is not intended to limit the invention to the precise form disclosed, and any such modifications, equivalents, and alternatives falling within the spirit and scope of the invention are intended to be included within the scope of the invention.
Claims (10)
1. A pseudomonas solanacearum (Pseudochrobactrum asaccharolyticum) strain not capable of degrading sugar, which is deposited in the China general microbiological culture Collection center, and has the deposit number: CGMCC No.28059.
2. A microbial agent comprising the pale bacillus oligosaccharide according to claim 1 as an active ingredient.
3. The fermentation method of the non-sugar-decomposing bacillus pseudolaris according to claim 1, comprising the steps of:
(1) Primary seed culture: inoculating the non-sugar-decomposing pseudomonas into a nutrient medium under the aseptic condition, and culturing for 12-24 hours at 25-35 ℃ and 150-300rpm to obtain a first-stage seed culture solution;
(2) Secondary seed culture: inoculating the primary seed culture solution into a nutrient medium according to an inoculum size of 1-10vol% under a sterile condition, and culturing for 12-24h at 25-35 ℃ and 150-300rpm to obtain a secondary seed culture solution;
(3) Fermentation: after the fermentation medium is sterilized, inoculating the secondary seed culture solution into the fermentation medium according to the inoculum size of 1-10vol%, controlling the temperature to be 25-35 ℃, the rotating speed to be 150-300rpm, fermenting under the condition that the aeration ratio is 1 (1-2), and stopping fermenting when dissolved oxygen starts to rise to obtain the fermentation liquid.
4. A fermentation process according to claim 3, wherein the nutrient medium has a composition of: 5-15g/L of peptone, 3-8g/L of yeast powder, 8-12g/L of sodium chloride, water as solvent and pH=6.5-7.5;
the composition of the fermentation medium is as follows: 5-15g/L of carbon source and 3-8g/L, PO of nitrogen source 4 3- 1-3g/L、K + 1-2g/L、Mg 2+ 0.05-0.10g/L、Ca 2+ 0.5-1.5g/L、Mn 2+ 0.05-0.1g/L, water as solvent, pH=6-8.
5. The fermentation process of claim 4, wherein the carbon source is selected from one or more of glucose, sucrose, starch, sodium acetate or sodium succinate;
the nitrogen source is selected from one or more of yeast extract powder, peptone, urea, ammonium sulfate or potassium nitrate.
6. A method for purifying waste water, comprising the step of inoculating the activated liquid of the sucrose-free pseudomonas solanacearum according to claim 1 or the microbial agent according to claim 2 into the waste water.
7. The method according to claim 6, wherein the concentration of sulfides in the wastewater is 400mg/L or less, preferably 300mg/L or less, most preferably 200mg/L or less.
8. The method according to claim 6 or 7, wherein the inoculation amount of the activation solution of the candida utilis insoluble in sugar or the microbial agent comprising the candida utilis insoluble in sugar is 100ppm or more, preferably 100-1000ppm.
9. The use of the non-saccharolytic pseudopallium of claim 1 and the microbial agent of claim 2 in the field of environmental remediation.
10. The use according to claim 9, characterized in that the candida albicans and the microbial agents comprising the candida albicans are used for degrading sulfides, preferably ferrous sulfide, H 2 S、HS - And S is 2- 。
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