CN110205356A - A kind of differentiated management method of the pharmacy waste water based on MIC toxicity detection technology - Google Patents

A kind of differentiated management method of the pharmacy waste water based on MIC toxicity detection technology Download PDF

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CN110205356A
CN110205356A CN201910403966.2A CN201910403966A CN110205356A CN 110205356 A CN110205356 A CN 110205356A CN 201910403966 A CN201910403966 A CN 201910403966A CN 110205356 A CN110205356 A CN 110205356A
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waste water
water
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CN110205356B (en
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苏悦
王开彬
杨宇斯
杜佳丽
夏雨
王伟
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Hangzhou Xiuchuan Technology Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • C12Q1/06—Quantitative determination
    • 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

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Abstract

The present invention relates to water treatment fields, disclose a kind of differentiated management method of pharmacy waste water based on MIC toxicity detection technology, comprising: 1) take each representative sub-thread waste water from enterprise's beginning of production, carry out water quality detection respectively；2) G is determined respectively+And G‑Middle abundance accounting is primary dominant bacteria；3) G is inquired respectively in prokaryotes classifieds website+、G‑In reference culture；4) bacterial strain is activated, is mixed to prepare seed liquor；5) by wastewater dilution at different dilutions, then after seed liquor is inoculated into wastewater medium hatching culture, in conjunction with visually observing and OD600Detect strain growth situation；6) four grades of waste water are defined；7) according to the water analysis of waste water and grade, shunting management is carried out to each stock waste water.The method of the present invention is based on bio-toxicity detection technique, can be realized enterprise wastewater toxicity personalization detection for pharmacy waste water, achievees the purpose that adaptation to local conditions.

Description

A kind of differentiated management method of the pharmacy waste water based on MIC toxicity detection technology
Technical field
The present invention relates to water treatment field more particularly to a kind of differences of the pharmacy waste water based on MIC toxicity detection technology Change management method.
Background technique
Pharmaceuticals industry waste water toxic pollutant is many kinds of, and many specific pollutants are after concentration reaches certain threshold value Inhibiting effect even catastrophic collapse can be caused to the biological treatment of waste water, such as heavy metal, antibiotic, cyanide, nitrobenzene. In enterprise's sewage daily management, the bio-toxicity that operation personnel needs to intake to each stock has certain understanding, in organic loading The quantitative management for reinforcing toxic waste on the basis of management, controls toxic wastewater, and system is avoided unexpected efficiency drop occur The case where low, Xie Xu, dead mud.
For a long time, the detection method of toxicity of water body is all established both at home and abroad, but is all concentrated mainly on river water and life Apply flexibly it is waterborne, and specifically for pharmaceutical chemical industry industry waste water carry out bio-toxicity detection means do not have been reported that also.And it is existing Detection method of toxicity, for example, toxicity detection instrument such as U.S. BECHMAN instrument company Microtox, SDI company Delta Tox Alert of Tox, Merck company, Skaler company, Holland Tox Tracer, be all based on this type of photobacteria G-Bacterial strain and establish, G can not be had both+And G-The toxic characteristic of both bacterial strains；And different enterprises has different lifes Change system, major microorganisms type is also different, does not have specific aim with unified detection method.For this reason, it is necessary to set up It is a kind of specifically in the waste water management system of pharmacy corporation.
Summary of the invention
In order to solve the above-mentioned technical problems, the present invention provides a kind of pharmacy waste waters based on MIC toxicity detection technology Differentiated management method.The method of the present invention is based on MIC (minimal inhibitory concentration) bio-toxicity detection technique, is trained by wastewater dilution It supports bacterium 18 to 24 hours, the minimum waste strength that can inhibit that environmental microorganism is grown in culture medium is found, therefore, it is determined that waste water To Ecotoxicology size.According to MIC experimental result, the present invention can be realized enterprise wastewater toxicity for pharmacy waste water Propertyization detection, achievees the purpose that adaptation to local conditions.The specific technical proposal of the invention is: a kind of based on MIC toxicity detection technology The differentiated management method of pharmacy waste water, comprising the following steps:
1) each representative sub-thread waste water is taken from enterprise's beginning of production, carries out water quality detection respectively.
2) rear three sections of progress activated sludge samplings carry out micro- life after mixing from before enterprise's original biochemical treatment system Object genome extracts, then analyzes the microorganism structure flora of sample, determines G respectively+And G-Middle abundance accounting is first Dominant bacteria, be denoted as G respectively+、G-(Gram is positive, negative), and the abundance ratio of the two is recorded, it is denoted as: m: n.
3) G is inquired respectively in prokaryotes classifieds website such as LPSN (http://www.bacterio.net/) etc.+、 G-In reference culture, be denoted as G respectively+A、G-B。
4) respectively to G+A、G-B bacterial strain is activated, to each strain growth to OD600When for 0.45-0.55, according to m: n Ratio mixing, is made seed liquor.
5) wastewater dilution to be measured is fabricated to the culture medium of gradient concentration at different dilutions；Seed liquor is inoculated into again It is used as test group in the culture medium of each gradient concentration, concurrently sets blank negative control group and blank positive controls；Hatching training After supporting, in conjunction with visually observing and OD600Detect strain growth situation.
6) according to the dilution of waste water and the upgrowth situation of bacterial strain, four grades of waste water are defined: nontoxic, less toxic, poisoning, High poison.
7) according to the water analysis of waste water and grade, shunting management is carried out to each stock waste water:
For high poison waste water, oxidation detoxification treatment is individually carried out or directly as dangerous waste processing；
For waste water of being poisoned: carrying out the materialized pretreatment of the advanced oxidation categories such as iron carbon, Fenton, biochemical system is entered after detoxification；
For less toxic waste water: this strand of waste water of institutionalization strict control enters the water of biochemical system, makes the CODcr tribute of less toxic part Ratio control is offered 50% hereinafter, otherwise taking materialized pretreatment means；
For nontoxic waste water: being directly entered biochemical treatment system.
The present invention has the advantages that seldom proposing to carry out Classification Management to waste water in existing Industrial Wastewater Treatment, generally not Classify, it is not fine accurate enough having classified, such as by survey BOD5, the fabulous waste water of biochemical can only be identified, The waste water of low toxicity, poisoning, high poison is acted on without differentiation, can only finally carry out materialized pretreatment together, so that having one Determine biochemical, originally can directly be degraded by the CODcr that biochemical degradation falls by materialization, occupies the resource at materializing strategy end, no It only results in processing cost to greatly improve, it is also possible to reduce the treatment effect of pretreatment section.And the method for the present invention is based on bio-toxicity Detection technique can be realized enterprise wastewater toxicity personalization detection for pharmacy waste water, carry out sophisticated category to waste water, and make Different processing modes so as to efficent use of resources, is greatly reduced cost, and excludes to bear pretreatment section treatment effect Face is rung.
The present invention makees above-mentioned the reason of being handled differently to different grades of waste water respectively and is: team of the present invention is actually grinding It finds during studying carefully: even if to poisoning, high poison waste water water in biochemical system is smaller to cause to whole system flora Irreversible damage seriously affects the treatment effeciency of biochemical treatment system, it is therefore desirable to carry out after independent detoxification again into biochemistry.And it is right The Biostatic of strain is located at the waste water in less toxic section, in the case that CODcr contribution rate is no more than 50% in biochemical system, Due to the diluting effect of nontoxic waste water, although having certain influence to microbial activity, be unlikely to make microflora degradation effect by compared with Big to influence, the further amount of inlet water that controls can reduce its threat to total system, and nontoxic waste water then can directly be tasted Biological treatment means are tried, expensive materializing strategy section pressure is reduced.
Preferably, the water quality detection includes following index: CODcr, NH in step 1)3- N, TN, TP, pH and TDS.
Preferably, the waste water is cephalo production line waste water.
Preferably, in step 4), the bacterial strain activation method are as follows:
A one end far from strain) is managed in freeze-drying to heat in alcolhol burner, drips sterile water, keeps freeze-drying Guan Yitou broken because of quenching It splits, completes open pipe.
B sterile water) is drawn with pasteur pipet, freeze-drying pipe pressure-vaccum dissolution bacterial strain powder repeatedly is added, obtain bacterium powder solution；
C bacterium powder solution) is drawn, drop one drips in the test tube for being pre-loaded with LB culture medium, completes inoculation；Each bacterial strain activates every time At least three pipe of inoculation, in order to avoid because accidentalia causes activation to fail.
D) inoculated test tube is sealed, is put into 33-37 DEG C of incubator 100-150rpm shaken cultivation activation 6-18h；It can See that culture medium obviously becomes cloudy.
E) with cannula type scene photometer read bacterial concentration, λ=600nm, as test tube bacterium solution OD=0.45-0.55, Growth logarithmic phase is arrived, is transferred in the conical flask of the LB culture medium equipped with dilution three times and expands culture, identical Under the conditions of culture to OD=0.45-0.55 when, activation is completed.
Preferably, step C) and E) in, inoculum concentration 0.8-1.2wt%.
Preferably, in step 5), the preparation side of the test group and blank negative control group, blank positive controls Method are as follows:
A) waste water to be measured is taken to adjust to neutrality, (if any suspended matter, first coagulation is heavy with 0.22 μm of filter filtration sterilization in desinfection chamber Shallow lake degerming again), and each gradient concentration is diluted to sterile water.
B) waste water of each gradient concentration and LB culture medium are mixed into test tube, each concentration makees four parallel pipes, separately If the test tube of multiple sterile water+LB culture mediums is bisected into two groups as blank positive control and blank negative control group.
C) in the test tube and blank positive controls activated seed liquor being inoculated into each test group with pipettor； There are a test tubes not to be inoculated with seed liquor in blank negative control group and the test group of each gradient concentration, and accesses sterile water and replace Generation.
Preferably, in step c), inoculum concentration 0.8-1.2wt%.
Preferably, waste water accounting is respectively after the dilution gradient concentration of the waste water is designed as dilution in step 5) 5%, 10%, 20%, 30%, 40% and 50%.
Each gradient at least three is parallel, needs to repeat test when parallel sample gap is larger and is confirmed.
Preferably, hatching condition of culture is the optimum growth temperature of bacterial strain, time 20-30h in step 5).
Preferably, the criterion of each grade of waste water is respectively as follows: in step 6)
Nontoxic: strain can normal growth under each dilution gradient；
Low toxicity: 40%≤minimal inhibitory concentration≤50%；
Poisoning: 10%≤minimal inhibitory concentration < 40%；
High poison: minimal inhibitory concentration < 10%.
Team of the present invention has summed up above-mentioned standard restriction, which is limited to reality after accumulating a large amount of practical experience The implementation of border of murder by poisoning identification in to(for) waste water is more accurate, to be conducive to select the corresponding most reasonable processing mode of waste water.
It is compared with the prior art, the beneficial effects of the present invention are:
1) present invention determines the standard reference of the enterprise wastewater by representing microbe species in analysis enterprise wastewater biochemical system Bacterial strain realizes the purpose that enterprise's toxicity detection is treated with a certain discrimination, so that detection is more targeted.
2) G is used simultaneously+And G-Two kinds of standard reference strains have taken into account two kinds of design features of microorganism, so that waste water is raw Object toxicity detection is more comprehensively accurate.
3) enterprise's raw wastewater is directlyed adopt as culture medium and carries out reference culture culture, is solved strain and is adapted in waste water Difficult problem.
4) it by carrying out grade delimitation to wastewater biological toxicity, solves the problems, such as that waste water is blindly handled, promotes enterprise real Existing science carries out water inlet management.
Specific embodiment
The present invention will be further described with reference to the examples below.
Total embodiment
A kind of differentiated management method of the pharmacy waste water based on MIC toxicity detection technology, comprising the following steps:
1) each representative sub-thread waste water is taken from enterprise's beginning of production, carries out water quality detection respectively, water quality detection includes following index: CODcr、NH3- N, TN, TP, pH and TDS.
CODcr measurement: it is measured using National Standard Method HJ828-2017, JC-101 type COD constent temperature heater；
NH3- N measurement: using even China Tech skill 6B-5C (V8) multi-parameter water quality analyzer measurement；
TN measurement: it is measured using National Standard Method HJ636-2012, ultraviolet-uisible spectrophotometer；
TP measurement: using even China Tech skill 6B-5C (V8) multi-parameter water quality analyzer measurement；
PH measurement: Shanghai thunder magnetic PHS-3S precision pH meter is used；
TDS measurement: it is measured using HJ/T 51-1999.
2) rear three sections of progress activated sludge samplings carry out micro- life after mixing from before enterprise's original biochemical treatment system Object genome extracts, then analyzes the microorganism structure flora of sample, determines G respectively+And G-Middle abundance accounting is first Dominant bacteria, be denoted as G respectively+、G-(Gram is positive, negative), and the abundance ratio of the two is recorded, it is denoted as: m: n.
3) G is inquired respectively in prokaryotes classifieds website such as LPSN (http://www.bacterio.net/)+、G- In reference culture, be denoted as G respectively+A、G-B。
4) respectively to G+A、G-B bacterial strain is activated:
A one end far from strain) is managed in freeze-drying to heat in alcolhol burner, drips sterile water, keeps freeze-drying Guan Yitou broken because of quenching It splits, completes open pipe.
B sterile water) is drawn with pasteur pipet, freeze-drying pipe pressure-vaccum dissolution bacterial strain powder repeatedly is added, obtain bacterium powder solution；
C bacterium powder solution) is drawn, drop one drips in the test tube for being pre-loaded with LB culture medium, completes inoculation, inoculum concentration 0.8- 1.2wt%；Each bacterial strain activates three pipe of at least inoculation every time, in order to avoid because accidentalia causes activation to fail.
D) inoculated test tube is sealed, is put into 33-37 DEG C of incubator 100-150rpm shaken cultivation activation 6-18h；It can See that culture medium obviously becomes cloudy.
E) with cannula type scene photometer read bacterial concentration, λ=600nm, as test tube bacterium solution OD=0.45-0.55, Growth logarithmic phase is arrived, is transferred in the conical flask of the LB culture medium equipped with dilution three times and expands culture, inoculum concentration When cultivated under the same conditions for 0.8-1.2wt% to OD=0.45-0.55, activation is completed.
Bacterium solution is mixed according to m: n ratio, seed liquor is made.
5) wastewater dilution to be measured is fabricated to the culture medium of gradient concentration at different dilutions；Seed liquor is inoculated into again It is used as test group in the culture medium of each gradient concentration, concurrently sets blank negative control group and blank positive controls；In bacterial strain Optimum growth temperature, after hatching culture 20-30h, in conjunction with visually observing and OD600Detect strain growth situation.
Preferably, test group and blank negative control group, blank positive controls the preparation method comprises the following steps:
A) waste water to be measured is taken to adjust to neutrality, (if any suspended matter, first coagulation is heavy with 0.22 μm of filter filtration sterilization in desinfection chamber Shallow lake degerming again), and each gradient concentration is diluted to sterile water.
B) waste water of each gradient concentration and LB culture medium are mixed into test tube, each concentration makees four parallel pipes, separately If the test tube of multiple sterile water+LB culture mediums is bisected into two groups as blank positive control and blank negative control group.
C) in the test tube and blank positive controls activated seed liquor being inoculated into each test group with pipettor； There are a test tubes not to be inoculated with seed liquor in blank negative control group and the test group of each gradient concentration, and accesses sterile water and replace Generation, access amount are 0.8-1.2wt%.
6) according to the dilution of waste water and the upgrowth situation of bacterial strain, four grades of waste water are defined: nontoxic, less toxic, poisoning, High poison.The criterion of each grade of waste water is respectively as follows:
Nontoxic: strain can normal growth under each dilution gradient；
Low toxicity: 40%≤minimal inhibitory concentration≤50%；
Poisoning: 10%≤minimal inhibitory concentration < 40%；
High poison: minimal inhibitory concentration < 10%.
7) according to the water analysis of waste water and grade, shunting management is carried out to each stock waste water:
For high poison waste water, oxidation detoxification treatment is individually carried out or directly as dangerous waste processing；
For waste water of being poisoned: carrying out the materialized pretreatment of the advanced oxidation categories such as iron carbon, Fenton, biochemical system is entered after detoxification；
For less toxic waste water: this strand of waste water of institutionalization strict control enters the water of biochemical system, makes the CODcr tribute of less toxic part Ratio control is offered 50% hereinafter, otherwise taking materialized pretreatment means；
For nontoxic waste water: being directly entered biochemical treatment system.
Embodiment 1
Zhejiang pharmaceutcal corporation, Ltd is one and specializes in cephalo-type, trains class medicine intermediate of muttering, bulk pharmaceutical chemicals, preparation production National new high-tech enterprise.8 strands of waste water that the present embodiment has specially taken its Cefaclor Lily synthesis technology to generate, number are 1-8。
Specific steps are as follows:
1) each representative 8 strands of waste water is taken from enterprise's beginning of production, carries out water quality detection respectively.
2) rear three sections of progress activated sludge samplings carry out micro- life after mixing from before enterprise's original biochemical treatment system Object genome extracts, then is analyzed (prior art) to the microorganism structure flora of sample, determines G respectively+And G-Middle abundance accounts for Than being denoted as G respectively for primary dominant bacteria+、G-(Gram is positive, negative), learns content highest in the waste water after analysis G+, G-Respectively Bacillus and Proteus, and the abundance ratio of two kinds of strains is 3: 1.2.
3) G is inquired respectively in prokaryotes classifieds website such as LPSN (http://www.bacterio.net/)+、G- In reference culture, be denoted as G respectively+A、G-B。G+A is Unite States Standard Culture Collection Center susceptibility bacterial strain Bacillus Subtilis ATCC6633, G-B are Proteus vulgaris ATCC 6380.
4) G+A, G-B bacterial strain are activated respectively:
A one end far from strain) is managed in freeze-drying to heat in alcolhol burner, dripping sterile water, makes to be lyophilized above Guan Yitou because rapid Open pipe is completed in cold and fragmentation.
B about 0.5ml sterile water) is drawn with pasteur pipet, and freeze-drying pipe pressure-vaccum dissolution strain powder repeatedly is added.
C bacterium powder solution) is drawn, drips (about 30 μ l) to being pre-loaded in the test tube of 3ml LB culture medium, that is, completes to connect Kind.Each strain activates three pipe of at least inoculation every time, in order to avoid because accidentalia causes activation to fail.
D) inoculated test tube is stoppered test tube plug, is put into 35 DEG C of incubator 120rpm shaken cultivations 12 hours and activates, it is seen that Culture medium obviously becomes cloudy.
E bacterial concentration (λ=600nm)) is read with cannula type scene photometer, as test tube bacterium solution OD=0.5 or so, i.e., To growth logarithmic phase, it is transferred in the conical flask of the LB culture medium equipped with dilution three times and expands culture (inoculum concentration 1%).When OD=0.5 or so is arrived in culture under the same conditions, activation is completed, and mixes bacterium solution by abundance ratio after activation, and obtained kind Sub- liquid.
5) by wastewater dilution to be measured at different dilutions (after dilution waste water accounting be respectively 5%, 10%, 20%, 30%, 40% and 50%), it is fabricated to the culture medium of gradient concentration；Seed liquor is inoculated into again in the culture medium of each gradient concentration as examination Group is tested, blank negative control group and blank positive controls are concurrently set；After bacterial strain optimum growth temperature, hatching culture for 24 hours, In conjunction with visually observing and OD600Detect strain growth situation.
Wherein, test group and blank negative control group, blank positive controls the preparation method comprises the following steps:
A) water sample to be measured is taken to adjust to neutrality, desinfection chamber with 0.22 μm of filter filtration sterilization (if any suspended matter, first coagulating sedimentation Degerming again), and each gradient concentration in 5) is diluted to sterile water.
B) the waste water 2.5ml of each dilution gradient and 0.5ml LB culture medium are mixed into test tube, each gradient does four Parallel pipe, the test tube for separately setting six 2.5ml sterile water+0.5ml LB culture mediums are bisected into two groups as blank positive control and sky White negative control group.
C) three test tubes and blank sun being inoculated into activated 30 μ l of bacterium solution with pipettor in each gradient test group In property control group；There are a test tubes not to be inoculated with bacterium solution as control feminine gender in blank negative control group and each gradient test group Control, and access the sterile water substitution of 30 μ l.
6) according to the dilution of waste water and the upgrowth situation of bacterial strain, four grades of waste water are defined: nontoxic, less toxic, poisoning, High poison.
7) according to the water analysis of waste water and grade, shunting management is carried out to each stock waste water.
Experimental result:
After the hatching culture of step 5), upgrowth situation under each concentration of all kinds of waste water is observed.Experimental result is as shown in table 1.
All kinds of waste water qualities of table 1 and MIC testing result
Note: ++: bacterial strain is obviously grown；+: the faint growth of bacterial strain；: bacterial strain is not grown；Wherein the 3rd group: the waste water is in each concentration Under, bacterial strain is not grown；2nd and 4 group: bacterial strain can be grown under each concentration.
Interpretation of result
The toxic degree of all kinds of waste water takes following criterion, as shown in table 2:
The toxic degree criterion of all kinds of waste water of table 2
Statistical analysis, the toxicity such as table 3 of all kinds of waste water in the present embodiment:
All kinds of wastewater toxicities of table 3 statistics
Toxicity Ratio Specific waste water (stock)
High poison 1/8=12.5% 3
Poisoning 1/8=12.5% 7
Low toxicity 4/8=50.0% 1、5、6、8
It is nontoxic 2/8=25.0% 4
Waste water treatment system water inlet guiding opinion
By table 1-3 interpretation of result, in 8 strands of waste water being investigated in addition to the 4th strand, hc effluent is belonged to, CODcr is in 40000- 100000mg/L.Wherein belong to high poison waste water for the 3rd strand, it is proposed that after carrying out detoxification treatment, then carry out subsequent biological treatment.
It is recommended that investigating noxious materials such as the 7th strand of waste water compositions such as raw material, solvent, product, by-product, controlled from source Toxic emissions processed.
It is recommended that directly carrying out biochemical treatment to 2,4 strands of waste water, 1,5,6,8 strand of waste water then gives up according to water is ascending one by one Water enters biochemical system, when certain strand of water into it is larger to the Inhibition of degradation of strain after biochemical system when it is then that this strand of waste water is individually pre- Biochemical system is entered back into after processing.
To keep enterprise wastewater management more systematic, more thorough, it is proposed that all strands of each workshop waste water is carried out MIC biology Toxicity detection.
Raw materials used in the present invention, equipment is unless otherwise noted the common raw material, equipment of this field；In the present invention Method therefor is unless otherwise noted the conventional method of this field.
The above is only presently preferred embodiments of the present invention, is not intended to limit the invention in any way, it is all according to the present invention Technical spirit any simple modification, change and equivalent transformation to the above embodiments, still fall within the technology of the present invention side The protection scope of case.

Claims (10)

1. a kind of differentiated management method of the pharmacy waste water based on MIC toxicity detection technology, it is characterised in that including following step It is rapid:
1) each representative sub-thread waste water is taken from enterprise's beginning of production, carries out water quality detection respectively；
2) rear three sections of progress activated sludge samplings carry out microorganism base after mixing from before enterprise's original biochemical treatment system Because of a group extraction, then the microorganism structure flora of sample is analyzed, determines G respectively+And G-Middle abundance accounting is primary excellent Gesture strain, is denoted as G respectively+、G-, and the abundance ratio of the two is recorded, it is denoted as: m:n；
3) G is inquired respectively in prokaryotes classifieds website+、G-In reference culture, be denoted as G respectively+A、G-B；
4) respectively to G+A、G-B bacterial strain is activated, to each strain growth to OD600When for 0.45-0.55, according to the ratio of m:n Seed liquor is made in mixing；
5) wastewater dilution to be measured is fabricated to the culture medium of gradient concentration at different dilutions；Seed liquor is inoculated into each ladder again It spends and is used as test group in the culture medium of concentration, concurrently set blank negative control group and blank positive controls；After hatching culture, In conjunction with visually observing and OD600Detect strain growth situation；
6) according to the dilution of waste water and the upgrowth situation of bacterial strain, four grades of waste water are defined: nontoxic, less toxic, poisoning, height Poison；
7) according to the water analysis of waste water and grade, shunting management is carried out to each stock waste water:
For high poison waste water, oxidation detoxification treatment is individually carried out or directly as dangerous waste processing；
For waste water of being poisoned: carrying out the materialized pretreatment of advanced oxidation class, biochemical system is entered after detoxification；
For less toxic waste water: this strand of waste water of institutionalization strict control enters the water of biochemical system, makes the CODcr tribute of less toxic part Ratio control is offered 50% hereinafter, otherwise taking materialized pretreatment means；
For nontoxic waste water: being directly entered biochemical treatment system.
2. the method as described in claim 1, which is characterized in that in step 1), the water quality detection includes following index: CODcr、NH3- N, TN, TP, pH and TDS.
3. the method as described in claim 1, which is characterized in that the waste water is cephalo production line waste water.
4. the method as described in claim 1, which is characterized in that in step 4), the bacterial strain activation method are as follows:
A one end far from strain) is managed in freeze-drying to heat in alcolhol burner, drips sterile water, keeps freeze-drying Guan Yitou broken because of quenching It splits, completes open pipe；
B sterile water) is drawn with pasteur pipet, freeze-drying pipe pressure-vaccum dissolution bacterial strain powder repeatedly is added, obtain bacterium powder solution；
C bacterium powder solution) is drawn, drop one drips in the test tube for being pre-loaded with LB culture medium, completes inoculation；
D) inoculated test tube is sealed, is put into 33-37 DEG C of incubator 100-150 rpm shaken cultivation activation 6-18h；
E bacterial concentration) is read with cannula type scene photometer, the nm of λ=600 arrives bacterium as test tube bacterium solution OD=0.45-0.55 Kind growth logarithmic phase is transferred to expand in the conical flask of the LB culture medium equipped with dilution three times and be cultivated, under the same conditions When cultivating OD=0.45-0.55, activation is completed.
5. method as claimed in claim 4, which is characterized in that step C) and E) in, inoculum concentration 0.8-1.2wt%.
6. the method as described in claim 1, which is characterized in that in step 5), the test group and blank negative control group, Blank positive controls the preparation method comprises the following steps:
A) it takes waste water to be measured to adjust to neutrality, in 0.22 μm of filter filtration sterilization of desinfection chamber, and is diluted to respectively with sterile water A gradient concentration；
B) waste water of each gradient concentration and LB culture medium are mixed into test tube, each concentration makees four parallel pipes, separately sets more The test tube of a sterile water+LB culture medium is bisected into two groups as blank positive control and blank negative control group；
C) in the test tube and blank positive controls activated seed liquor being inoculated into each test group with pipettor；Blank There are a test tubes not to be inoculated with seed liquor in negative control group and the test group of each gradient concentration, and accesses sterile water substitution.
7. method as claimed in claim 6, which is characterized in that in step c), inoculum concentration 0.8-1.2wt%.
8. method as described in claim 1 or 6, which is characterized in that in step 5), the dilution gradient concentration of the waste water is designed It is respectively 5%, 10%, 20%, 30%, 40% and 50% for waste water accounting after dilution.
9. the method as described in claim 1, which is characterized in that in step 5), hatching condition of culture is the most suitable growth of bacterial strain Temperature, time 20-30h.
10. the method as described in claim 1, which is characterized in that in step 6), the criterion of each grade of waste water is distinguished Are as follows:
Nontoxic: strain can normal growth under each dilution gradient；
Low toxicity: 40%≤minimal inhibitory concentration≤50%；
Poisoning: 10%≤minimal inhibitory concentration < 40%；
High poison: minimal inhibitory concentration < 10%.
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CN113023891A (en) * 2021-04-07 2021-06-25 知和环保科技有限公司 Front-end wastewater toxicity early warning and disposal device and application method thereof
CN114426934A (en) * 2021-08-20 2022-05-03 杭州秀川科技有限公司 Lactobacillus plantarum for source wastewater biotoxicity detection and application thereof

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CN108459146A (en) * 2018-01-10 2018-08-28 中冶华天工程技术有限公司 Assess method of the waste water to water treatment micro-organism toxicity

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CN108459146A (en) * 2018-01-10 2018-08-28 中冶华天工程技术有限公司 Assess method of the waste water to water treatment micro-organism toxicity

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Publication number Priority date Publication date Assignee Title
CN113023891A (en) * 2021-04-07 2021-06-25 知和环保科技有限公司 Front-end wastewater toxicity early warning and disposal device and application method thereof
CN114426934A (en) * 2021-08-20 2022-05-03 杭州秀川科技有限公司 Lactobacillus plantarum for source wastewater biotoxicity detection and application thereof
CN114426934B (en) * 2021-08-20 2023-07-11 杭州秀川科技有限公司 Lactobacillus plantarum for detecting biotoxicity of source wastewater and application thereof

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