CN117586929B - 降解菌株及其在养殖废水处理中的应用 - Google Patents
降解菌株及其在养殖废水处理中的应用 Download PDFInfo
- Publication number
- CN117586929B CN117586929B CN202410079846.2A CN202410079846A CN117586929B CN 117586929 B CN117586929 B CN 117586929B CN 202410079846 A CN202410079846 A CN 202410079846A CN 117586929 B CN117586929 B CN 117586929B
- Authority
- CN
- China
- Prior art keywords
- strain
- degradation
- benzimidazole
- fenbendazole
- microbial inoculum
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 230000015556 catabolic process Effects 0.000 title claims abstract description 39
- 238000006731 degradation reaction Methods 0.000 title claims abstract description 39
- 238000004065 wastewater treatment Methods 0.000 title description 3
- HYZJCKYKOHLVJF-UHFFFAOYSA-N 1H-benzimidazole Chemical compound C1=CC=C2NC=NC2=C1 HYZJCKYKOHLVJF-UHFFFAOYSA-N 0.000 claims abstract description 24
- 229960002669 albendazole Drugs 0.000 claims abstract description 24
- HXHWSAZORRCQMX-UHFFFAOYSA-N albendazole Chemical compound CCCSC1=CC=C2NC(NC(=O)OC)=NC2=C1 HXHWSAZORRCQMX-UHFFFAOYSA-N 0.000 claims abstract description 24
- 239000002351 wastewater Substances 0.000 claims abstract description 24
- 230000000507 anthelmentic effect Effects 0.000 claims abstract description 22
- 239000002068 microbial inoculum Substances 0.000 claims abstract description 18
- 229960005473 fenbendazole Drugs 0.000 claims abstract description 17
- IRHZVMHXVHSMKB-UHFFFAOYSA-N fenbendazole Chemical compound [CH]1C2=NC(NC(=O)OC)=NC2=CC=C1SC1=CC=CC=C1 IRHZVMHXVHSMKB-UHFFFAOYSA-N 0.000 claims abstract description 17
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 7
- 238000009629 microbiological culture Methods 0.000 claims abstract description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 28
- 239000001963 growth medium Substances 0.000 claims description 16
- 230000001580 bacterial effect Effects 0.000 claims description 14
- 239000011780 sodium chloride Substances 0.000 claims description 14
- 239000002609 medium Substances 0.000 claims description 13
- 230000000593 degrading effect Effects 0.000 claims description 9
- 229940124339 anthelmintic agent Drugs 0.000 claims description 8
- 239000000921 anthelmintic agent Substances 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 8
- 229960003439 mebendazole Drugs 0.000 claims description 7
- 238000009360 aquaculture Methods 0.000 claims description 6
- 244000144974 aquaculture Species 0.000 claims description 6
- 238000000855 fermentation Methods 0.000 claims description 6
- 230000004151 fermentation Effects 0.000 claims description 6
- BAXLBXFAUKGCDY-UHFFFAOYSA-N mebendazole Chemical compound [CH]1C2=NC(NC(=O)OC)=NC2=CC=C1C(=O)C1=CC=CC=C1 BAXLBXFAUKGCDY-UHFFFAOYSA-N 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 5
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- 239000007787 solid Substances 0.000 claims description 4
- 229910021536 Zeolite Inorganic materials 0.000 claims description 3
- HNPSIPDUKPIQMN-UHFFFAOYSA-N dioxosilane;oxo(oxoalumanyloxy)alumane Chemical compound O=[Si]=O.O=[Al]O[Al]=O HNPSIPDUKPIQMN-UHFFFAOYSA-N 0.000 claims description 3
- 239000010457 zeolite Substances 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 2
- 230000000813 microbial effect Effects 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 21
- 230000012010 growth Effects 0.000 abstract description 10
- 244000052616 bacterial pathogen Species 0.000 abstract description 5
- 239000000243 solution Substances 0.000 description 10
- 239000003814 drug Substances 0.000 description 6
- 239000002054 inoculum Substances 0.000 description 6
- 239000000356 contaminant Substances 0.000 description 5
- 229910017053 inorganic salt Inorganic materials 0.000 description 5
- 241000238631 Hexapoda Species 0.000 description 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 4
- 229940041514 candida albicans extract Drugs 0.000 description 4
- 239000008367 deionised water Substances 0.000 description 4
- 229910021641 deionized water Inorganic materials 0.000 description 4
- 210000003608 fece Anatomy 0.000 description 4
- 239000010871 livestock manure Substances 0.000 description 4
- 239000012138 yeast extract Substances 0.000 description 4
- 108020004465 16S ribosomal RNA Proteins 0.000 description 3
- 241000607528 Aeromonas hydrophila Species 0.000 description 3
- 241000607473 Edwardsiella <enterobacteria> Species 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- 102000029749 Microtubule Human genes 0.000 description 3
- 108091022875 Microtubule Proteins 0.000 description 3
- 241000607594 Vibrio alginolyticus Species 0.000 description 3
- 241000607272 Vibrio parahaemolyticus Species 0.000 description 3
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000003344 environmental pollutant Substances 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 230000000749 insecticidal effect Effects 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 210000004688 microtubule Anatomy 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 231100000719 pollutant Toxicity 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- 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 2
- 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 2
- 241000282414 Homo sapiens Species 0.000 description 2
- 229930195725 Mannitol Natural products 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 244000046052 Phaseolus vulgaris Species 0.000 description 2
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000010227 cup method (microbiological evaluation) Methods 0.000 description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 2
- 231100000053 low toxicity Toxicity 0.000 description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 2
- 235000019341 magnesium sulphate Nutrition 0.000 description 2
- 239000000594 mannitol Substances 0.000 description 2
- 235000010355 mannitol Nutrition 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 239000002957 persistent organic pollutant Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 239000010802 sludge Substances 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- TWFZGCMQGLPBSX-UHFFFAOYSA-N Carbendazim Natural products C1=CC=C2NC(NC(=O)OC)=NC2=C1 TWFZGCMQGLPBSX-UHFFFAOYSA-N 0.000 description 1
- 239000004215 Carbon black (E152) Substances 0.000 description 1
- 239000012880 LB liquid culture medium Substances 0.000 description 1
- HRRBJVNMSRJFHQ-UHFFFAOYSA-N Naftopidil Chemical compound COC1=CC=CC=C1N1CCN(CC(O)COC=2C3=CC=CC=C3C=CC=2)CC1 HRRBJVNMSRJFHQ-UHFFFAOYSA-N 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 206010043275 Teratogenicity Diseases 0.000 description 1
- 102000004243 Tubulin Human genes 0.000 description 1
- 108090000704 Tubulin Proteins 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 244000000054 animal parasite Species 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- YTLQFZVCLXFFRK-UHFFFAOYSA-N bendazol Chemical compound N=1C2=CC=CC=C2NC=1CC1=CC=CC=C1 YTLQFZVCLXFFRK-UHFFFAOYSA-N 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- JNPZQRQPIHJYNM-UHFFFAOYSA-N carbendazim Chemical compound C1=C[CH]C2=NC(NC(=O)OC)=NC2=C1 JNPZQRQPIHJYNM-UHFFFAOYSA-N 0.000 description 1
- 239000006013 carbendazim Substances 0.000 description 1
- 231100000315 carcinogenic Toxicity 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 239000000645 desinfectant Substances 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000037149 energy metabolism Effects 0.000 description 1
- 230000002550 fecal effect Effects 0.000 description 1
- 229960004500 flubendazole Drugs 0.000 description 1
- CPEUVMUXAHMANV-UHFFFAOYSA-N flubendazole Chemical compound C1=C2NC(NC(=O)OC)=NC2=CC=C1C(=O)C1=CC=C(F)C=C1 CPEUVMUXAHMANV-UHFFFAOYSA-N 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000003673 groundwater Substances 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 150000002430 hydrocarbons Chemical class 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 231100000219 mutagenic Toxicity 0.000 description 1
- 230000003505 mutagenic effect Effects 0.000 description 1
- 229950005705 naftopidil Drugs 0.000 description 1
- 239000008239 natural water Substances 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 238000011392 neighbor-joining method Methods 0.000 description 1
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 229960004454 oxfendazole Drugs 0.000 description 1
- BEZZFPOZAYTVHN-UHFFFAOYSA-N oxfendazole Chemical compound C=1C=C2NC(NC(=O)OC)=NC2=CC=1S(=O)C1=CC=CC=C1 BEZZFPOZAYTVHN-UHFFFAOYSA-N 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 230000036284 oxygen consumption Effects 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 238000012257 pre-denaturation Methods 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 238000004153 renaturation Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000010865 sewage Substances 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 231100000378 teratogenic Toxicity 0.000 description 1
- 230000003390 teratogenic effect Effects 0.000 description 1
- 231100000211 teratogenicity Toxicity 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000000844 transformation Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/22—Bacillus
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P1/00—Disinfectants; Antimicrobial compounds or mixtures thereof
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/38—Organic compounds containing nitrogen
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/40—Organic compounds containing sulfur
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/20—Nature of the water, waste water, sewage or sludge to be treated from animal husbandry
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
-
- 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
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- Pest Control & Pesticides (AREA)
- General Engineering & Computer Science (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Environmental Sciences (AREA)
- Plant Pathology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Agronomy & Crop Science (AREA)
- Water Supply & Treatment (AREA)
- Environmental & Geological Engineering (AREA)
- Dentistry (AREA)
- Hydrology & Water Resources (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明公开了一株降解菌株及其在养殖废水处理中的应用，降解菌株为贝莱斯芽孢杆菌(Bacillus velezensis），命名为WR‑4，于2023年12月18日保藏于中国微生物菌种保藏管理委员会普通微生物中心，菌种保藏编号为：CGMCC No.29354。本发明的菌株可有效降解养殖废水中的苯并咪唑类抗蠕虫药，对甲苯咪唑、芬苯达唑、阿苯达唑或丁苯咪唑的降解率可达85%以上，同时菌剂对COD、TP和TN也具有较好的降解效果，还可抑制病原菌的生长，在养殖尾水中具有良好应用前景。
Description
技术领域
本发明属于微生物技术领域，具体涉及一株降解菌株及其在养殖废水处理中的应用。
背景技术
养殖废水中的污染物主要含有有机物、微生物和有毒有害物。有机污染物主要是养殖场粪污中含有的碳氢化合物、含氮、含磷有机物和未被消化的营养物质，排放进入自然水体后，可使水体固体悬浮物、化学耗氧量、生化需氧量升高。当超量的有机污染物进入水体后，超过其通过稀释、沉淀、吸附、分解、降解等作用的自净能力时，水质便会恶化，这种水体很难再净化和恢复生机。微生物污染主要是养殖场粪污中含有大量的病原微生物，它们随粪进入水体后，以水为媒介进行传播和扩散，造成某些疫病的传播和扩散，危害人和动物的健康并带来经济损失。有毒有害污染物主要是指饲料中的抗生素、药物及猪场用的消毒剂等，随粪污排入水体后会对土壤和地下水造成污染。
其中，苯并咪唑类药物（BMZs）是一中常用的广谱、高效、低毒的抗蠕虫药，主要的药物有甲苯咪唑、芬苯达唑、阿苯达唑、丁苯咪唑、奥芬达唑、奥苯达唑、三氯苯咪唑、坎苯哒唑、氟苯达唑、鲁苯达唑、萘托比明、多菌灵等。因其具有驱虫谱广、驱虫效果好、毒性低，还有一定的杀灭幼虫和虫卵的作用，被广泛应用于动物寄生虫的治疗，同时还可作为杀霉菌剂应用于农产品的储存和运输。苯并咪唑类药物抗虫作用的机理主要是与虫体的微管蛋白选择性结合，抑制微管增长，引起微管的损伤，阻止微管组装的聚合，从而影响虫体的消化和营养吸收，干扰虫体的能量代谢。
研究发现，苯并咪唑类抗蠕虫药抑制细胞活性的作用是其具有潜在的致突变性和致畸性。当人们长期食用此类药物残留的动物性食品时，这些残留物便会在人体聚集，严重可能会致癌、致畸、致突变，最终危害到人类和动物的健康。
发明内容
为解决现有技术的不足，本发明的目的在于提供一株能够高效降解养殖废水中苯并咪唑类抗蠕虫药、COD、TP、TN等浓度水平的菌株，并可抑制相关病原菌生长。
为了实现上述目标，本发明采用如下的技术方案：
一株降解菌株，降解菌株为贝莱斯芽孢杆菌(Bacillus velezensis），命名为WR-4，于2023年12月18日保藏于中国微生物菌种保藏管理委员会普通微生物中心，菌种保藏编号为：CGMCC No.29354。
上述降解菌株降解养殖废水中苯并咪唑类抗蠕虫药的应用，苯并咪唑类抗蠕虫药为甲苯咪唑、芬苯达唑、阿苯达唑或丁苯咪唑中的一种；养殖废水中苯并咪唑类抗蠕虫药的浓度为50~200mg/L，NaCl浓度为0.5%~1.5%，pH为6~8，温度为10~30℃。
利用上述降解菌株制备的菌剂。
一种上述菌剂降解养殖废水中苯并咪唑类抗蠕虫药的应用，苯并咪唑类抗蠕虫药为甲苯咪唑、芬苯达唑、阿苯达唑或丁苯咪唑中的一种；养殖废水中苯并咪唑类抗蠕虫药的浓度为50~200mg/L，NaCl浓度为0.5%~1.5%，pH为6~8，温度为10~30℃。
一种利用上述菌剂降解养殖废水中苯并咪唑类抗蠕虫药的方法，具体步骤为：将菌剂按照质量体积比为0.5~1.5g/L的投加量投加至养殖废水中，搅拌均匀，进行降解。
一种上述菌剂的制备方法，包括以下具体步骤：
S1、将菌株WR-4接入LB培养基中培养，得到种子液；
S2、将种子液接入发酵培养基中扩大培养，得到菌液；
S3、将菌液与沸石粉混合，烘干得到固体菌剂。
优选地，前述步骤S1中，LB培养基的组分为：酵母提取物 5g、蛋白胨 10g、NaCl 5g和水1L 。
优选地，前述步骤S2中，发酵培养基的组分为：葡萄糖15～20g/L、豆粕粉15～20g/L、酵母膏4～8g/L、甘露醇5～8g/L、磷酸氢二钾1～2g/L和硫酸镁0.5～1g/L；种子液的接种量为2%~ 5%。
优选地，前述病原菌为副溶血弧菌、嗜水气单胞菌、溶藻弧菌和爱德华氏菌中的一种。
本发明的有益之处在于：本发明筛选出一株贝莱斯芽孢杆菌(Bacillus velezensisWR-4)，该菌株可有效降解养殖废水中的苯并咪唑类抗蠕虫药，对甲苯咪唑、芬苯达唑、阿苯达唑或丁苯咪唑的降解率可达85%以上，可高效去除废水中的COD、TP和TN，还可抑制病原菌的生长，在养殖尾水中具有良好应用前景。
附图说明
图1为菌株WR-4的菌落形态图；
图2为菌株WR-4的系统发育树；
图3为菌株WR-4在不同温度培养基中的OD生长曲线；
图4为菌株在不同盐度下的OD600值；
图5是本发明的菌株WR-4对苯并咪唑类抗蠕虫药的降解效果图；
图6是不同污染物初始浓度条件下WR-4对阿苯达唑的降解效果图；
图7是不同pH条件下WR-4对阿苯达唑的降解效果图；
图8为不同温度条件下WR-4对阿苯达唑的降解效果图；
图9为菌剂对养殖废水的降解效果图。
实施方式
以下结合附图和具体实施例对本发明作具体的介绍。
实施例1 菌株WR-4的分离筛选及鉴定：
从连云港市赣榆区青口镇养殖尾水污泥获取样本，经PBS缓冲液梯度稀释后，充分摇匀后吸取50 μL稀释液，分别涂布于培养基平板上，在30℃下好氧培养24-48 h。待菌落形成后挑取单菌落再次平板划线分离，得到菌种种类单一的菌落，共计20株单菌。将各菌分别接种于配有100mg/L阿苯达唑浓度的无机盐培养基中，测定各菌降解阿笨达唑的效果，结果显示菌株WR-4对阿苯达唑表现出优异降解特性，其菌落形态图如图1所示，表面凸起呈乳白色、不透明、圆形、边缘不规则、表面褶皱。
本发明测定了菌株的16S rDNA序列，其序列表详见SEQ ID NO：1。细菌扩增的引物采用细菌通用引物。正向引物为Eubac27F:(5-AGAGTTTGATC-CTGGCTCAG-3) ，其序列表详见SEQ ID NO：2，反向引物为Eubac1492R:(5- GGTTACCTTGTTACGACTT-3) ，其序列表详见SEQID NO：3，将25 μL 2×Mix Taq与正反向引物各2 μL、菌液2 μL混匀后，加水至50 μL体系后进行PCR扩增。PCR反应程序为94℃预变性4 min；94℃变性30 s；55℃复性1 min；72℃延伸1.5 min，共30个循环；72℃延伸10 min。利用1%的琼脂糖凝胶电泳检测PCR产物后送至测序公司进行测序，对菌株16S rDNA序列结果采用BLAST方法在NCBI数据库搜索比对分析。将菌株WR-4及其相近菌株的16S rDNA序列通过Mega(Version7.0)软件进行多序列比对，并采用Neighbor-joining方法构建系统进化树，如图2。
经鉴定，该菌为贝莱斯芽孢杆菌，并命名为Bacillus velezensis WR-4。
上述菌株于2023年12月18日保藏于中国微生物菌种保藏管理委员会普通微生物中心，其分类命名为贝莱斯芽孢杆菌(Bacillus velezensis)，菌株名为WR-4，保藏地址为：北京市朝阳区北辰西路1号院3号，菌种保藏号为：CGMCC No.29354。
实施例2 菌剂的制备：
将菌株Bacillus velezensis WR-4接入100 mL LB培养基中，于30℃，180 rpm摇床中震荡培养24h得到种子液，之后将种子液按体积比1%的接种量接入发酵培养基的发酵罐中进行扩大培养，培养至109 CFU/mL浓度时，4000 rpm离心收集菌泥；将菌泥与沸石粉按1：3体积混合，35℃低温烘干得到固体菌剂。
其中，LB培养基的组分为：酵母提取物 5g、蛋白胨 10g、NaCl 5g和水1L，培养条件为：温度 35℃，时间 24h，pH为6～8。
发酵培养基的组分为：葡萄糖15～20g/L、豆粕粉15～20g/L、酵母膏4～8g/L、甘露醇5～8g/L、磷酸氢二钾1～2g/L和硫酸镁0.5～1g/L，培养条件为：温度 35℃，时间36 h，pH为6～8。
实施例3 不同温度下菌株WR-4的生长能力测定：
将实施例1获得的菌株以体积比1％的接种量接种至10 mL的LB液体培养基中活化，在温度分别为10℃、20℃、30℃、40℃、50℃的条件下，按2%的接种量将菌株接种于pH为7的LB培养基中、180 rpm摇床中连续震荡培养48h，间隔6h测定不同温度下培养基中的OD600值，结果如图3所示。
由图3可知，菌株WR-4在30℃的温度下较早进入生长对数期，因此最佳温度为30℃，且其温度耐受性为10~40℃。
实施例4 不同盐度下菌株WR-4的生长性能测定：
在LB液体培养基基础上，通过添加或减少无水NaCl，使培养基中NaCl浓度分别为0、0.5%、1%、1.5%、2.0%、2.5%、3.0%、3.5%。将1%的种子液分别接种到100mL的上述不同盐度的LB液体培养基中，30℃、180rpm恒温震荡培养培养20h，测定其吸光度OD600值，如图4所示。
由图4可知，菌株WR-4在0.5%~1.5%的NaCl溶液下生长效果最好，当NaCl溶液浓度大于1.5%时，生长能力随着盐浓度的上升而下降。
实施例5 菌株WR-4对苯并咪唑类抗蠕虫药的降解效果测定：
将菌株WR-4以2%的接种量分别接种到含甲苯咪唑、芬苯达唑、阿苯达唑和丁苯咪唑的无机盐培养基中，培养基中各污染物的浓度均为100 mg/L，培养基组分为NH4NO3 0.5g、MgSO4 0.33g、NaCl 0.88g、FeSO4·7H2O 1.2g、Na2HPO4•12H2O 0.6g、MnSO4·H2O 0.56g、(NH4)2SO41.2g，去离子水补足至1L，pH 7.0，然后将装有污染物的培养基分别置于温度30℃、转速180rpm的条件下培养48h后，测定降解各污染物效果，结果如图5所示。
由图5可以看出，菌株WR-4对甲苯咪唑、芬苯达唑、阿苯达唑或丁苯咪唑的降解率在48h均可达到85%以上，降解效果优异。
实施例6 污染物初始浓度对菌株WR-4降解效果的影响：
在阿苯达唑初始浓度分别为10mg/L、20mg/L、50mg/L、100mg/L、200mg/L和300mg/L的条件下，按2%的接种量，分别将菌种WR-4接入无机盐培养基中，培养基组分为NH4NO3 0.5g、MgSO4 0.33g、NaCl 0.88g、FeSO4·7H2O 1.2g、Na2HPO4•12H2O 0.6g、MnSO4·H2O0.56g、(NH4)2SO41.2g，去离子水补足至1L，pH 7.0，然后将装有污染物的培养基分别置于温度30℃、转速180rpm的条件下培养48h后，降解效果如图6所示。
由图6可以看出，阿苯达唑的浓度为50~200mg/L时，48h内降解率均能达到80%，72h后，降解率可达98%以上。
实施例7 pH对菌株WR-4降解效果的影响：
在pH分别为5、6、7、8、9、10的条件下，按2%的接种量，将菌株WR-4接种至含100 mg/L阿苯达唑的无机盐培养基中，培养基组分为NH4NO3 0.5g、MgSO4 0.33g、NaCl 0.88g、FeSO4·7H2O 1.2g、Na2HPO4•12H2O 0.6g、MnSO4·H2O 0.56g、(NH4)2SO41.2g，去离子水补足至1L，置于相同温度30℃、相同转速180 rpm的条件下培养48h，降解效果结果如图7所示。
由图7可知，pH在6~8范围内，菌株WR-4对阿苯达唑的降解率能达到85%以上，效果显著；pH为10条件下，降解率也有45%，说明该菌株在碱性较强的环境中也具有一定的降解效果。
实施例8 不同温度下菌株WR-4对阿苯达唑的降解效果测定：
在温度分别为5℃、8℃、10℃、20℃、30℃、40℃、50℃的条件下，按2%的接种量，将菌株WR-4接种至含100 mg/L阿苯达唑的无机盐培养基中，培养基组分为NH4NO3 0.5g、MgSO4 0.33g、NaCl 0.88g、FeSO4·7H2O 1.2g、Na2HPO4•12H2O 0.6g、MnSO4·H2O 0.56g、(NH4)2SO41.2g，去离子水补足至1L，置于相同转速180rpm，pH 7.0，的条件下培养48h，结果如图8所示。
由图8可知，8~40℃温度下，菌株WR-4对阿苯达唑的降解率可达到60%以上；而10~30℃温度下，降解率可达到88%以上。
实施例9 菌剂对养殖废水的处理效果：
取5m3某养殖场的养殖废水，其中阿苯达唑、COD、TP和氨氮的浓度分别为100mg/L、200mg/L、5mg/L和50mg/L，将5kg菌剂均匀投加至养殖废水中，并搅拌均匀。于30℃降解48h，降解效果如图9所示。
由图9可知，菌剂对阿苯达唑、COD、TP、氨氮的降解效率分别为 89.5%、91.7%、54.78%、68.46%，此时菌株活菌数为106CFU/mL左右。以上实验数据表明，菌剂不仅可以降解养殖废水中的苯并咪唑类抗蠕虫药，且可有效降解养殖废水中的COD、TP和氨氮，在处理养殖废水领域中具有良好的应用前景。
实施例10 抑菌实验（牛津杯法）：
将养殖尾水中常见的副溶血弧菌、嗜水气单胞菌、溶藻弧菌、爱德华氏菌活化备用，采用牛津杯法测定菌株WR-4的抑菌能力，滴加磷酸盐缓冲液作为阴性对照，通过抑菌圈大小来表示抑制效果。结果显示WR-4对副溶血弧菌、嗜水气单胞菌、溶藻弧菌、爱德华氏菌的抑制效果分别为18mm±0.34、15mm±0.17、16.7mm±0.28、13mm±0.31。
通过该实验可知，WR-4能够有效抑制养殖尾水中常见的病原菌的生长。
以上显示和描述了本发明的基本原理、主要特征和优点。本行业的技术人员应该了解，上述实施例不以任何形式限制本发明，凡采用等同替换或等效变换的方式所获得的技术方案，均落在本发明的保护范围内。
Claims (6)
1.一株降解菌株，其特征在于，所述降解菌株为贝莱斯芽孢杆菌(Bacillus velezensis），命名为WR-4，于2023年12月18日保藏于中国微生物菌种保藏管理委员会普通微生物中心，菌种保藏编号为：CGMCC No.29354。
2.权利要求1所述的降解菌株在降解养殖废水中苯并咪唑类抗蠕虫药中的应用，其特征在于，所述苯并咪唑类抗蠕虫药为甲苯咪唑、芬苯达唑、阿苯达唑或丁苯咪唑中的一种；所述养殖废水中苯并咪唑类抗蠕虫药的浓度为50~200mg/L，NaCl浓度为0.5%~1.5%，pH为6~8，温度为10~30℃。
3.利用权利要求1所述的降解菌株制备的菌剂。
4.一种权利要求3所述的菌剂在降解养殖废水中苯并咪唑类抗蠕虫药中的应用，其特征在于，所述苯并咪唑类抗蠕虫药为甲苯咪唑、芬苯达唑、阿苯达唑或丁苯咪唑中的一种；所述养殖废水中苯并咪唑类抗蠕虫药的浓度为50~200mg/L，NaCl浓度为0.5%~1.5%，pH为6~8，温度为10~30℃。
5.一种权利要求3所述的菌剂降解养殖废水中苯并咪唑类抗蠕虫药的方法，其特征在于，将菌剂按照质量体积比为0.5~1.5g/L的投加量投加至养殖废水中，搅拌均匀，进行降解。
6.一种权利要求3所述的菌剂的制备方法，其特征在于，包括以下具体步骤：
S1、将菌株WR-4接入LB培养基中培养，得到种子液；
S2、将种子液接入发酵培养基中扩大培养，得到菌液；
S3、将菌液与沸石粉混合，烘干得到固体菌剂。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410079846.2A CN117586929B (zh) | 2024-01-19 | 2024-01-19 | 降解菌株及其在养殖废水处理中的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410079846.2A CN117586929B (zh) | 2024-01-19 | 2024-01-19 | 降解菌株及其在养殖废水处理中的应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN117586929A CN117586929A (zh) | 2024-02-23 |
| CN117586929B true CN117586929B (zh) | 2024-03-26 |
Family
ID=89913808
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202410079846.2A Active CN117586929B (zh) | 2024-01-19 | 2024-01-19 | 降解菌株及其在养殖废水处理中的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117586929B (zh) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117736938B (zh) * | 2024-02-07 | 2024-05-17 | 南京万瑞环境科技有限公司 | 一种微生物复合菌群及其应用 |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102286412A (zh) * | 2011-08-30 | 2011-12-21 | 新疆农业科学院微生物应用研究所 | 一种贝莱斯芽孢杆菌及其在番茄枯萎病拮抗中的应用 |
| CN115353986A (zh) * | 2022-01-28 | 2022-11-18 | 齐齐哈尔大学 | 用于处理养猪废水的贝莱斯芽孢杆菌菌株wb株及其应用 |
| CN116790439A (zh) * | 2023-07-13 | 2023-09-22 | 江苏理工学院 | 一株贝莱斯芽孢杆菌及其在降解废水cod的应用 |
-
2024
- 2024-01-19 CN CN202410079846.2A patent/CN117586929B/zh active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102286412A (zh) * | 2011-08-30 | 2011-12-21 | 新疆农业科学院微生物应用研究所 | 一种贝莱斯芽孢杆菌及其在番茄枯萎病拮抗中的应用 |
| CN115353986A (zh) * | 2022-01-28 | 2022-11-18 | 齐齐哈尔大学 | 用于处理养猪废水的贝莱斯芽孢杆菌菌株wb株及其应用 |
| CN116790439A (zh) * | 2023-07-13 | 2023-09-22 | 江苏理工学院 | 一株贝莱斯芽孢杆菌及其在降解废水cod的应用 |
Non-Patent Citations (1)
| Title |
|---|
| Microbial degradation of the benzimidazole fungicide carbendazim by Bacillus velezensis HY‑3479;Suyoung Song 等;International Microbiology;20230915;第1-9页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN117586929A (zh) | 2024-02-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109182193B (zh) | 微生物菌剂及其制备方法和应用 | |
| CN117586929B (zh) | 降解菌株及其在养殖废水处理中的应用 | |
| CN101591069B (zh) | 受污染湖泊水体的处理方法 | |
| Li et al. | Cometabolic biodegradation of antibiotics by ammonia oxidizing microorganisms during wastewater treatment processes | |
| CN109762773B (zh) | 一种树状类芽孢杆菌、固定化菌剂及其应用 | |
| CN109182178B (zh) | 一株具有铬耐受和Cr(VI)去除能力的菌株及其在原位修复中轻度铬污染土壤中的应用 | |
| CN110819556B (zh) | 一种根瘤菌及其菌剂和应用 | |
| CN116640702A (zh) | 一种用于处理养殖废水的微生物菌剂的制备方法和应用 | |
| CN113215033B (zh) | 一种磺胺类抗生素降解菌及其应用 | |
| CN117603888B (zh) | 一株蜡样芽孢杆菌及其在养殖尾水处理中的应用 | |
| CN110791444A (zh) | 一种施氏假单胞菌、采用该施氏假单胞菌制得的复合菌剂及该复合菌剂的应用 | |
| CN104862260A (zh) | 一株具有好氧反硝化能力的节杆菌及其应用 | |
| CN107460148B (zh) | 一种枯草芽孢杆菌菌粉的制备方法及应用 | |
| CN100579920C (zh) | 一种生活污水处理方法 | |
| EP1210407B1 (en) | Bacterial consortium ebc1000 and a method using the bacterial consortium ebc1000 for remedying biologically recalcitrant toxic chemicals contained in industrial wastewater, waste materials and soils | |
| CN113957004B (zh) | 一株金黄杆菌及在制备盐生植物附生修复养护菌剂中的应用 | |
| CN108277175A (zh) | 2，4二硝基甲苯磺酸盐高效降解菌株Microbacterium sp.X3及其应用 | |
| CN110791450A (zh) | 一株能够降解多种β-内酰胺类抗生素的菌株Am101及其应用 | |
| CN115386520B (zh) | 一株嗜吡啶红球菌rl-gz01菌株及其应用 | |
| CN115125178B (zh) | 一株具有四环素类抗生素降解功能的类芽孢杆菌、方法及应用 | |
| CN109652328B (zh) | 一种复合微生物活菌制剂及其在高浓度养猪废水中的应用 | |
| CN111172144A (zh) | 一种高效降解四环素类抗生素的微生物菌剂及其在土壤污染修复中的应用 | |
| CN101993838A (zh) | 具有氯苯胺降解能力的代尔夫特菌h1及其应用 | |
| KR20140077645A (ko) | 수질 및 저질 개선용 미생물제제 제형 | |
| Phatthongkleang et al. | The Efficiency of Bacillus spp. to Remove Ammonia in Shrimp Aquaculture |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |