CN105969705A - Stabilizing agent for selenium-contaminated soil repairing bacteria - Google Patents

Stabilizing agent for selenium-contaminated soil repairing bacteria Download PDF

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Publication number
CN105969705A
CN105969705A CN201610612556.5A CN201610612556A CN105969705A CN 105969705 A CN105969705 A CN 105969705A CN 201610612556 A CN201610612556 A CN 201610612556A CN 105969705 A CN105969705 A CN 105969705A
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China
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selenium
parts
soil
sodium
stabilizer
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CN201610612556.5A
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袁志辉
周海燕
雍婕
程益
肖红仕
吴永尧
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Hunan University of Science and Engineering
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Hunan Agricultural University
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • 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
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
    • B09C—RECLAMATION OF CONTAMINATED SOIL
    • B09C1/00—Reclamation of contaminated soil
    • B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes

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  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
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  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Wood Science & Technology (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Environmental & Geological Engineering (AREA)
  • Virology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Medicinal Chemistry (AREA)
  • Soil Sciences (AREA)
  • Mycology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Processing Of Solid Wastes (AREA)

Abstract

The invention discloses a stabilizing agent for selenium-contaminated soil repairing bacteria and belongs to the technical field of environmental microorganisms. The stabilizing agent is prepared from 10-20 parts of glycerin, 10-20 parts of saccharose, 5-10 parts of glucose, 2-6 parts of sodium succinate, 2-10 parts of sodium nitrate, 1-8 parts of sodium nitrite, 5-10 parts of an ultraviolet light absorber, 1-10 parts of disodium hydrogen phosphate, 1-10 parts of sodium dihydrogen phosphate and 5-10 parts of calcium peroxide. The stabilizing agent can increase the content of oxygen in soil, can enhance the survival ability, the adaptive ability to the environment and the repair ability of bacteria in the selenium-contaminated soil repairing process, and has good practical application prospects.

Description

A kind of selenium contamination soil remediation antibacterial stabilizer
Technical field
The invention belongs to technical field of environmental microorganism, relate to a kind of selenium contamination soil remediation antibacterial stabilizer.
Background technology
Some area such as enshi and Ankang of China then belongs to high selenium area, its Selenium in Soils content severe overweight and cause the generation of the situations such as selenosis such as depilation, anemia.Further, since the development of selenium-rich industry, in the production process of selenium-enriched crop, constantly in soil, apply selenium preparation so that many agricultural land soils are changed into selenium from the state that the most scarce selenium or selenium are appropriate and exceed standard or even the situation of selenium contamination.Therefore, need case above to carry out selenium contamination repairing and treating.The major way of Selenium in Soil pollution control at present is phytoremediation, but plant growing cycle is long, and results plant is difficult to the defects such as process.Recent study finds, sodium selenite the strongest for soil poisoning and sodium selenate can be reduced to the elemental nano-selenium that toxicity is relatively low by many microorganisms, become the state that plant is difficult to absorb, reduce the selenium content of plant, be by the environmental friendliness of selenium contamination soil remediation, method with low cost.
During utilizing microorganism to carry out selenium contamination environmental improvement, there are distinct issues is exactly that the condition of selenium contamination environment is sufficiently complex, and oxygen content is low in soil environment, long by the ultraviolet irradiation time in sunlight, used microorganism vitality wherein is declined can not survive the most completely, the microorganism remediation efficiency ultimately resulting in selenium contamination environment declines, and can not repair the most completely.The open of practical technique of this type of difficult problem is not the most overcome to report with in document in the market.Therefore, as developed a kind of selenium contamination soil remediation antibacterial stabilizer, improve function strain stability in complex environment, survival ability and remove selenium ability, then can effectively solve to repair the problem of strain environment-adapting ability difference, effectively play microorganism and except selenium advantage and solve the problem that selenium contamination environmental microorganism remediation efficiency is low.
Summary of the invention
It is an object of the invention to provide a kind of selenium contamination soil remediation antibacterial stabilizer.This stabilizer can increase oxygen content in soil, improve function strain stability in complex environment, survival ability and remove selenium ability, can effectively solve to repair the problem of strain environment-adapting ability difference, effectively play microorganism and except selenium advantage and solve the problem that selenium contamination environmental microorganism remediation efficiency is low.
The selenium contamination soil remediation antibacterial stabilizer that the present invention provides, form as follows: glycerol 10-20 part, sucrose 10-20 part, glucose 5-10 part, sodium succinate 2-6 part, sodium nitrate 2-10 part, sodium nitrite 1-8 part, UV absorbent 5-10 part, disodium hydrogen phosphate 1-10 part, sodium dihydrogen phosphate 1-10 part, calper calcium peroxide 5-10 part.
UV absorbent in stabilizer is phenyl salicylate, 2-(2-hydroxyl-5-aminomethyl phenyl) BTA, oxybenzoic acid phenyl ester, 2,4-dihydroxy benaophenonel, ESCALOL 567, Octabenzone, 2-(2 '-hydroxyl-3 ', 5 '-two tertiary phenyl)-5-chlorination benzotriazole, one or more in single benzoic acid resorcinol.
The selenium contamination soil remediation antibacterial stabilizer that the present invention provides, can make the soil oxygen content at 30 cm increase to close with air oxygen content i.e. about 20% from 12%；Quantity after antibacterial 24 h being applied in soil reaches 1.2 × 105-1.5×107Individual/g soil, much larger than the soil remediation bacterial number being not added with stabilizer；Selenite content in contaminated soil is the most relatively not added with the rehabilitating soil of stabilizer and declines faster.Therefore, the selenium contamination soil remediation antibacterial stabilizer that the present invention provides can be effectively improved repairs the survival ability of antibacterial and except selenium efficiency, has preferable actual application prospect.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is described in further details, but is not limited to the scope of the present invention.Embodiment is implemented under premised on technical solution of the present invention, gives detailed embodiment and concrete operating process.In following embodiment, method therefor is conventional method if no special instructions.
Embodiment 1:
The preparation of selenium contamination soil remediation antibacterial stabilizer:
Glycerol 10-20 part, sucrose 10-20 part, glucose 5-10 part, sodium succinate 2-6 part, sodium nitrate 2-10 part, sodium nitrite 1-8 part, UV absorbent 5-10 part, disodium hydrogen phosphate 1-10 part, sodium dihydrogen phosphate 1-10 part, calper calcium peroxide 5-10 part, the material of above number is sufficiently mixed uniformly.
UV absorbent in stabilizer is phenyl salicylate, 2-(2-hydroxyl-5-aminomethyl phenyl) BTA, oxybenzoic acid phenyl ester, 2,4-dihydroxy benaophenonel, ESCALOL 567, Octabenzone, 2-(2 '-hydroxyl-3 ', 5 '-two tertiary phenyl)-5-chlorination benzotriazole, one or more in single benzoic acid resorcinol.
Embodiment 2:
Selenium contamination soil remediation bacteria screening is prepared with mycopowder:
Repair bacteria screening: with containing 0.2 mM Na2SeO3LB flat board be isolation medium, the pedotheque of collection is diluted to 10-1To 10-8Suspension, takes acceptable diluent degree and is coated on separation flat board, cultivate 1-15d for 25 DEG C.Select and can reduce Na2SeO3Produce the object bacteria that the antibacterial (preliminary basis for estimation is red colonies) of red elemental selenium is studied as next step.By primary dcreening operation obtain antibacterial renewed vaccination to containing with without Na2SeO3LB flat board on further determine whether that there is reduction Na2SeO3Produce the ability of red elemental selenium, record form preservation.At 0-100 Mm Na2SeO3Carry out Selenium endurance mensuration on the LB flat board of Concentraton gradient, select the strongest bacterial strain of Selenium endurance as repairing bacterial strain.
Prepared by mycopowder: preparation LB flat board, and activation repairing bacterial strain of ruling cultivates 24 h at 37 DEG C；Single colony inoculation of growth on activation flat board, in LB fluid medium, is cultivated 24 h as seed culture fluid at 37 DEG C；The seed liquor prepared is inoculated in the LB fluid medium containing 1 mM sodium selenite by the inoculum concentration of 3%, 37 DEG C, 120 rpm cultivation 48h.Skim milk after adding 20% sterilizing in the bacteria suspension obtained, freezing at-20 ~-50 DEG C, it is dried under vacuum acquisition mycopowder.
Application example one:
The stabilizer utilizing preparation repairs selenium contamination soil with repairing antibacterial mycopowder laboratory test, and detection wherein repairs bacteria living quantity and remediation efficiency, and its process and result are as follows:
A. high selenium zone, dam, autonomous prefecture fish pond of bestowing favour from Hubei Province selects 4 places to gather high-Se soils, removes the root, stem and leaf of crop and weeds etc., suitably pulverizes into little soil block or soil particle, gather and the process time is on July 6th, 2015；
B. the stabilizer of preparation is mixed homogeneously in the ratio of 20:1 with repairing antibacterial mycopowder；
C. the mixture in B is sufficiently mixed with selenium contamination pedotheque with the ratio of 20g/kg soil；
D. being spilled into certain water in the B pedotheque mixed, amount is advisable with ground moistening；
E. the soil handled well in D is loaded in plastic tub, places 48 h under room temperature state；
The most separately set the comparison being not added with stabilizer: in addition to as adding stabilizer, the same A-D of remaining process；
G. use heavy metal kind and content in GB GB 15618-1995 detection initial soil sample, use grandson to give the emerald green accelerated solvent extraction-Solid-Phase Extraction-high performance liquid chromatography-tandem mass (ASE-SPE-HPLC-MS/MS) waiting (2013) to introduce and analyze kind and the content of antibiotic in method detection pedotheque.Show after detection, gather the Cd in the high-Se soil on dam, autonomous prefecture fish pond of bestowing favour from Hubei Province2+、Ni2+、Pb2+And Co2+Exceeding national standard, tetracycline, ampicillin, Study on Determination of Gentamycin are all higher than 50 μ g/kg；
H. use Holtzclaw(1987) etc. introduction method detection Hubei Province bestow favour dam, autonomous prefecture fish pond untreated high-Se soil in+4 valency selenium average contents be 86.21 mg/kg, after processing 48 h, in soil ,+4 valency selenium average contents are reduced to 5.1 mg/kg, and compareing+4 average Se contents of valency in soil after processing 48 h is 53.6 mg/kg；
I. after stabilizer treatment 24 h is added in the detection of employing fluorescence microscope counting method, the reparation bacterial cell par in soil is 1.5 × 107Individual/g soil, the reparation bacterial cell par in comparison soil is 1.1 × 102Individual/g soil；
J. utilizing the detection of oxygen in soil content meter to add after stabilizer treatment 24 h oxygen average content in soil is 25.6%, and the oxygen average content compareed in soil after 24 h is 17.2%.
Application example two:
The stabilizer utilizing preparation repairs selenium contamination soil on the spot with repairing antibacterial mycopowder outdoor, and detection wherein repairs bacteria living quantity and remediation efficiency, and its process and result are as follows:
A. high selenium zone, dam, autonomous prefecture fish pond of bestowing favour in Hubei Province selects smooth plot, and area is about 10 m × 10 m, and process group and matched group respectively set 3 repetition, and the root, stem and leaf of removal crop and weeds etc., test period is on July 6th, 2015；
B. the stabilizer of preparation is mixed homogeneously in the ratio of 20:1 with repairing antibacterial mycopowder；
C., before turning in farmland, the mixture in B is spread in the farmland that Se content exceeds standard with the ratio uniform of 10kg/ mu, is turned in farmland according to a conventional method；
The most separately set the comparison being not added with stabilizer: in addition to as adding stabilizer, the same A-C of remaining process；
E. use GB GB 15618-1995 to detect heavy metal kind and content in original agricultural land soil sample, use grandson to give the emerald green accelerated solvent extraction-Solid-Phase Extraction-high performance liquid chromatography-tandem mass (ASE-SPE-HPLC-MS/MS) waiting (2013) and analyze kind and the content of antibiotic in method detection pedotheque.Show after detection, the Cr in test agricultural land soil2+、Cd2+、Ni2+、Pb2+And Co2+Exceed national standard, be not detected by antibiotic；
F. use Holtzclaw(1987) etc. introduction method detection test soil in+4 valency selenium average contents be 77.5 mg/kg, after processing 48 h, in soil ,+4 valency selenium average contents are reduced to 6.7 mg/kg, and in comparison soil ,+4 valency Se contents are 48.9 mg/kg；
G. after stabilizer treatment 24 h is added in the detection of employing fluorescence microscope counting method, the reparation bacterial cell par in soil is 1.2 × 105Individual/g soil, the reparation bacterial cell par in comparison soil is 1.2 × 102Individual/g soil；
H. utilizing the detection of oxygen in soil content meter to add after stabilizer treatment 24 h oxygen average content in 30 cm depths soil is 20.1%, and the oxygen average content compareed in 30 cm depths soil is 11%.

Claims (3)

1. a selenium contamination soil remediation antibacterial stabilizer.
2. the stabilizer described in claim 1 consists of: glycerol 10-20 part, sucrose 10-20 part, glucose 5-10 part, sodium succinate 2-6 part, sodium nitrate 2-10 part, sodium nitrite 1-8 part, UV absorbent 5-10 part, disodium hydrogen phosphate 1-10 part, sodium dihydrogen phosphate 1-10 part, calper calcium peroxide 5-10 part.
3. the UV absorbent described in claim 2 is phenyl salicylate, 2-(2-hydroxyl-5-aminomethyl phenyl) BTA, oxybenzoic acid phenyl ester, 2,4-dihydroxy benaophenonel, ESCALOL 567, Octabenzone, 2-(2 '-hydroxyl-3 ', 5 '-two tertiary phenyl)-5-chlorination benzotriazole, one or more mixing in single benzoic acid resorcinol.
CN201610612556.5A 2016-07-30 2016-07-30 Stabilizing agent for selenium-contaminated soil repairing bacteria Pending CN105969705A (en)

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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103952363A (en) * 2014-05-16 2014-07-30 华中农业大学 Bacillus ES2-45 capable of reducing selenite to produce nano-selenium and application thereof
CN104004520A (en) * 2014-05-07 2014-08-27 上海纺织建筑设计研究院 Arsenic-polluted place soil restoration stabilizer
CN104117534A (en) * 2013-04-25 2014-10-29 成都新朝阳作物科学有限公司 Combined application of zinc-selenium agricultural preparation and soil heavy metal repairing agent
CN105132314A (en) * 2015-08-31 2015-12-09 苏州硒谷科技有限公司 Brevibacterium frigoritolerans YLX-3 and application thereof
CN105176534A (en) * 2015-06-25 2015-12-23 青岛恩高运动控制技术有限公司 Material for soil remediation
CN105583217A (en) * 2015-12-23 2016-05-18 江苏盖亚环境工程有限公司 Conditioner for repairing heavy metal contaminated soil and preparing method for conditioner

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104117534A (en) * 2013-04-25 2014-10-29 成都新朝阳作物科学有限公司 Combined application of zinc-selenium agricultural preparation and soil heavy metal repairing agent
CN104004520A (en) * 2014-05-07 2014-08-27 上海纺织建筑设计研究院 Arsenic-polluted place soil restoration stabilizer
CN103952363A (en) * 2014-05-16 2014-07-30 华中农业大学 Bacillus ES2-45 capable of reducing selenite to produce nano-selenium and application thereof
CN105176534A (en) * 2015-06-25 2015-12-23 青岛恩高运动控制技术有限公司 Material for soil remediation
CN105132314A (en) * 2015-08-31 2015-12-09 苏州硒谷科技有限公司 Brevibacterium frigoritolerans YLX-3 and application thereof
CN105583217A (en) * 2015-12-23 2016-05-18 江苏盖亚环境工程有限公司 Conditioner for repairing heavy metal contaminated soil and preparing method for conditioner

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
利峰 等: "土壤硒污染的生物修复技术", 《广东微量元素科学》 *
朱维琴 等: "铁、锰等金属元素的微生物还原及其在环境生物修复中的意义", 《应用生态学报》 *

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Effective date of registration: 20190429

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Application publication date: 20160928