CN105986000A - Method for detecting biotoxicity of pollutants by virtue of mixed bacterium bio-sensor - Google Patents
Method for detecting biotoxicity of pollutants by virtue of mixed bacterium bio-sensor Download PDFInfo
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Abstract
The invention discloses a method for detecting biotoxicity of pollutants by virtue of a mixed bacterium bio-sensor. The method comprises the following steps: 1) preparing microbial liquid which consists of saccharomycetes liquid, escherichia coli liquid and bacillus subtilis liquid; 2) uniformly mixing the microbial liquid with a mixed sol, and coating an obtained mixture on flexible thin films, so that a microbial thin films are prepared; 3) assembling the microbial thin films on electrodes, so that the mixed bacterium bio-sensor is manufactured; 4) detecting a to-be-detected sample; and 5) evaluating the biotoxicity of the to-be-detected sample. The mixed bacterium bio-sensor can be used for evaluating single toxicity and joint toxicity of heavy metals, phenol and/or chemicals. The method has the characteristic of being convenient and rapid to detect; and meanwhile, the limitation that a single microbe can cause differences on sensitivity to toxic substances. The method not only improves the sensitivity of comprehensive toxicity analysis on a combined pollution system of a water body, but also reflects the actual situations on the toxicity of the pollutants.
Description
Technical field
The present invention relates to bio-toxicity detection field, more particularly to by fungus, gram negative bacteria and leather
The mixed vaccine biosensor that the mixed vaccine of Lan Shi positive bacteria composition is prepared as challenge organisms, to ring
Environment pollution thing carries out bio-toxicity detection.
Background technology
Along with fast development and the raising of people's living standard of industrial and agricultural production, synthetic chemical substance
Kind and consumption sharply increase, pesticide, chemical fertilizer, heavy metal, industrial wastewater and house refuse etc. are a large amount of
Enter environment, environment is caused severe contamination.Therefore in detection environment, the bio-toxicity of pollutant is being sentenced
The bio-safety degree in abscission ring border, environmental improvement and agricultural planting secure context suffer from important effect.
There is now the method utilizing single microorganism detection environmental contaminants bio-toxicity, but utilize single
When one microbial cell carries out bio-toxicity detection, it is vulnerable to the impact of microbe species, because the most of the same race
The sensitivity of the microbe toxicity of class is different, so toxicity assessment based on single bacterial strain has
The limitation of pollutant authentic virulence effect can not be objectively responded.It addition, existing activated sludge process evaluation
Bio-toxicity has certain limitation, because different location, the activated sludge bacteria group composition of different time collection
Differ greatly so that toxicity detection result does not has comparative.
It addition, in natural environment, pollute and not only caused by a kind of material, there is two or more chemistry
During material, the bio-toxicity between different material can produce synergism, i.e. can cause independent with each material
Diverse toxic reaction during effect.Up to now, the detection of most of bio-toxicities is both for single
The experiment of pollutant, existing water standard is also that the toxicity test according to single metal determines, it is impossible to
It is effectively reflected the collaborative poisonous effect of multiple noxious substance.Therefore two or more chemicals is studied
Joint toxic effect is increasingly becoming the important directions of environmental toxicology research, and it can be the formulation of environmental standard
More reliable foundation is provided with ecological risk assessment.
Accordingly, it is desirable to provide a kind of method detecting environmental contaminants, it can detect environmental contaminants
Bio-toxicity, can avoid the single microorganism limitation to the sensitivity of toxicant simultaneously, moreover it is possible to visitor
See the toxicity synergism reflecting multiple toxicant.
Summary of the invention
It is an object of the present invention to provide a kind of mixed vaccine biosensor detection pollutant bio-toxicity
Method, comprise the steps:
1) preparing microbial inoculum, it includes yeast liquid, Escherichia coli bacteria liquid and bacillus subtilis bacterium
Liquid；
2) it is coated on fexible film after described microbial inoculum being mixed with mixed sols, prepares microorganism
Thin film；
3) described microbial thin film is assembled on electrode, i.e. prepares mixed vaccine biosensor；
4) detection testing sample；
5) bio-toxicity of sample to be tested is evaluated.
The method wherein preparing microbial inoculum is:
1) microbial inoculant is cultivated to exponential phase in culture medium；
2) thalline is collected, it is thus achieved that bacterial sediment；
3) washing bacterial sediment；
4) disperse bacterial sediment with dispersion liquid, make microorganism dispersion liquid；
5) it is mixed in proportion different types of microorganism dispersion liquid, makes microbial inoculum.
In said method, what the culture medium of cultivating microorganism was well known in the art is suitable to growth of microorganism
Culture medium, the such as culture medium of escherichia coli inoculation are that LB culture medium, SOB culture medium or SOC cultivate
Base etc., preferably LB culture medium；The culture medium of described bacillus subtilis be beef-protein medium or
LBG culture medium, preferably beef-protein medium；Described saccharomycetic culture medium is that YPD cultivates
Base, YPED culture medium or YPEG culture medium etc., preferably YPED culture medium.
Further, the condition of culture of microorganism is also to it is known in the art that to include but not limited to, large intestine
The condition of culture of bacillus is 37 DEG C and cultivates 16-24h in LB culture medium；The cultivation bar of bacillus subtilis
Part is 37 DEG C and cultivates 16-24 hour in beef-protein medium；Saccharomycetic condition of culture is 30 DEG C
Cultivate 24-30 hour in YPED culture medium.
In the above-mentioned methods, it is preferable that collect the thalline cultivated by centrifuging, to obtain bacterial sediment.
The washing of bacterial sediment can use the buffer of routine, follow-up to remove possible impact in yeast culture
The material of test result.Such as phosphate buffer, Tris-Hcl buffer, HEPEs buffer, boric acid-boron
Sand buffer or sodium chloride solution.Preferably, described wash solution is sodium chloride solution, the PBS of 0.85%
Solution, TBS solution or TBE solution.
Preferably, described dispersion liquid is that aqueous solution, sodium chloride solution or culture medium, preferably sodium chloride are molten
Liquid, the sodium chloride solution of more preferably 0.85%.
Escherichia coli in microbial inoculum: bacillus subtilis: yeast is 1-3:1-3:1；Preferably, greatly
Enterobacteria: bacillus subtilis: yeast is 2:2:1.
Described mixed sols is polyvinyl alcohol (PVA) colloidal sol, polyvinyl alcohol-4-ethylpyridine graft polymers
Colloidal sol, polyvinyl alcohol-sodium sulfate colloidal sol or polyvinyl alcohol-alginate mixed sols, preferably polyvinyl alcohol
-alginate mixed sols.
Described fexible film is that polyurethane film, polydimethylsiloxanefilm film, polrvinyl chloride are thin
Film or pet film, preferably pet film.
Described electrode is platinum electrode, glass-carbon electrode, diamond electrode, preferably platinum electrode.
Described mixed vaccine biosensor is placed in 4 DEG C of preservations, and the holding time is 2-6 hour.
The method of described sample to be tested detection comprises the steps:
1) breathing medium is added；
2) apply voltage, and detect electric current.
3) add benzoquinone solution, and detect electric current, it is thus achieved that the first current intensity；
4) sample to be tested is added；
5) detection electric current, it is thus achieved that the second current intensity；
Described breathing medium includes sodium lactate, succinic acid, glucose and sodium chloride, preferably 10mM breast
Acid sodium, 10mM sodium succinate, 10mM glucose and 0.85% sodium chloride.
The voltage applied is 0.2V-0.7V (vs.Ag/AgCl), preferably 0.3V (vs.Ag/AgCl).
Described benzoquinone solution ultimate density is 0.1mM-1mM, preferably 0.4mM.
Evaluate the bio-toxicity of sample to be tested with suppression ratio, the computing formula of suppression ratio is as follows:
Suppression ratio (%)=(1-I2/I1) × 100%
Wherein, I1It is the first current intensity, I2It is the second current intensity.
Corresponding suppression ratio is can get, with the concentration of sample to be tested by the sample to be tested of detection variable concentrations
Carrying out curve fitting suppression ratio, suppression ratio is that the concentration of sample to be tested when 50% is defined as this and treats test sample
This IC50.And IC50 is calculated as 1 toxic unit (TU) of measured matter.
Cleaning Principle of the present invention is shown in Fig. 1.
Beneficial effects of the present invention is as follows:
The present invention, using mixed bacteria liquid as challenge organisms, prepares mixed vaccine biosensor, can be used for commenting
The single toxicity of valency heavy metal, phenol and/or pesticide and joint toxicity.Detection has fast and convenient feature,
Can avoid single culture that poisonous substance sensitivity is had discrepant restriction simultaneously.Not only improve the compound dirt of water body
The sensitivity that dye system comprehensive toxicity is analyzed, and the truth of pollutant toxicity can be reflected.
Accompanying drawing explanation
Below in conjunction with the accompanying drawings the detailed description of the invention of the present invention is described in further detail.
Fig. 1 is illustrated based on the ultimate principle figure of mixed vaccine biosensor detection noxious substance bio-toxicity.
Fig. 2 illustrates that mixed vaccine biosensor detects Cu2+The result of bio-toxicity.
Fig. 3 illustrates that mixed vaccine biosensor detects Cd2+The result of bio-toxicity.
Fig. 4 illustrates the result of mixed vaccine biosensor detection DCP bio-toxicity.
Fig. 5 illustrates the result of mixed vaccine biosensor detection orthene bio-toxicity.
Fig. 6 illustrates the result of mixed vaccine biosensor detection ametryn bio-toxicity.
Fig. 7 illustrates the result of mixed vaccine biosensor detection actual waste water bio-toxicity.
Detailed description of the invention
In order to be illustrated more clearly that the present invention, below in conjunction with preferred embodiments and drawings, the present invention is done into one
The explanation of step.Parts similar in accompanying drawing are indicated with identical reference.Those skilled in the art
Should be appreciated that following specifically described content is illustrative and be not restrictive, should not limit with this
Protection scope of the present invention.
Embodiment 1
The preparation of culture medium:
Culture medium (YEPD) compound method of saccharomyces cerevisiae is as follows: take 1g yeast extract and 2g peptone is molten
In 90ml ultra-pure water, take 1g glucose and be dissolved in 10ml ultra-pure water, in 120 DEG C of high-pressure sterilizing pots
Being sterilized separately 20min, after natural cooling, mixing can use.
Colibacillary culture medium (LB) compound method is as follows: take 0.5g Carnis Bovis seu Bubali cream, 1g peptone and
0.5g sodium chloride is dissolved in 100mL ultra-pure water, regulates pH to 7.0-7.4, in high pressure steam sterilization 120
DEG C sterilizing 20min, can use after natural cooling.
The culture medium configuration of bacillus subtilis is as follows: take 1g peptone, 0.3g beef extract and 0.5g
Sodium chloride is dissolved in 100mL ultra-pure water, regulation pH to 7.0, sterilizing in high-pressure steam sterilizing pan, from
So cooling.
The cultivation of microorganism:
To YPED inoculation of medium saccharomyces cerevisiae (S288C), cultivate 24h 30 DEG C of waters bath with thermostatic control,
Shaking table speed 200rpm；
To LB culture medium inoculated escherichia coli (ATCC 25922), condition of culture is 37 DEG C of water bath with thermostatic control trainings
Support 16h, shaking table speed 180rpm；
Bacillus subtilis (bacterium number: 1.1086), condition of culture is inoculated in the cultivation of bacillus subtilis
Be condition be that 24h, shaking table speed 180rpm are cultivated in 37 DEG C of waters bath with thermostatic control；
Will yeast culture to exponential phase.
Microorganism collection
Saccharomycetic collection: will cultivate the yeast centrifugation of gained, rotating speed is 10000rpm, centrifugal
10min, cleans the bacterial sediment of centrifugal gained twice with PBS, the bacterial sediment weight after then cleaning
Newly it is suspended in PBS, obtains certain density bacterium solution stand-by；
Escherichia coli and the collection of bacillus subtilis:
The rotating speed that centrifugation for the first time is used is 6000rpm, and the time is 6min.Subsequently with 0.85%
Bacterial sediment is cleaned twice by sodium chloride solution, and the rotating speed that twice cleaning uses is 5000rpm, and the time is
5min.Finally it is distributed to obtaining bacterial sediment after cleaning in 0.85% sodium chloride solution, obtains thalline and divide
Dissipate liquid.
Bacterium solution mixes:
Three kinds of thalline are hybridly prepared into mixed bacteria liquid, and mixed proportion is escherichia coli: bacillus subtilis:
Yeast=2:2:1.
The preparation of biofilm:
Take 100 μ l 10% (w/v) PVA colloidal sols (saponification number 98%, the degree of polymerization 2400), the 1.0M of 10 μ l
Sodium sulfate, 40 μ l 0.8% (w/v) sodium alginates and 50 μ l mixed vaccine suspension mix homogeneously, by gained
To mixed liquor be spun on pet film, be coated with the poly-to benzene of mixed vaccine
Naphthalate film is immersed in 2% (w/v) CaCl2In, under the conditions of 20-25 DEG C, keep 15-20min,
I.e. obtain disposable microbial thin film after drying, put into cold preservation in 4 DEG C of refrigerators stand-by.
The detection method of bio-toxicity:
At ambient temperature, biomembrane is fixed on platinum electrode surface, is assembled into mixed vaccine biosensor,
Adding the respiratory substrate of 10ml pH=7.0 in mixed vaccine biosensor, described breathing medium contains
10mM sodium lactate, 10mM sodium succinate, 10mM glucose and 0.85% sodium chloride, apply 0.3V
(vs.Ag/AgCl) voltage, uses chronoamperometry that size of current is carried out real-time monitoring；Treat body
System adds 100 μ l benzoquinone solution after stablizing 5min, and wherein the ultimate density of benzoquinone is 0.4mM, treats electric current
Signal stabilization, records the first current value (I1), it is subsequently adding poisonous substance, detects and record the second current value (I2)。
Utilize suppression ratio to calculate bio-toxicity, and judge the power of bio-toxicity according to IC50.IC50 is counted
1 toxic unit (TU) for measured matter.
The detection method of joint-biotoxicity:
Two kinds of identical for toxic unit poisonous substances are mixed and made into mixing sample by 1:1, according to described bio-toxicity
Detection method detect the suppression ratio of this mixing sample, and draw concentration-suppression ratio curve, it is thus achieved that this mixing
Actual measurement IC50 (the IC50 of sampleobs).By comparing expection IC50 value (IC50exp) and measured value IC50
(IC50obs) size judge the type of joint toxicity.Wherein expect that the computational methods of IC50exp are:
The IC50 value of two kinds of single poisonous substances is added.Work as IC50exp>IC50obsThe just biological poison of two kinds of poisonous substances of explanation
Property has antagonism；Expection IC50exp<IC50obsIllustrate that the bio-toxicity of two kinds of poisonous substances has collaborative work
With；IC50exp=IC50obs, illustrate that the bio-toxicity of two kinds of poisonous substances has summation action.Antagonism is connection
Close the summation of the bio-toxicity produced when bio-toxicity acts solely on body less than each chemical substance, be i.e.
A certain chemical substance can weaken the bio-toxicity of another chemical substance；Synergism is then the biological poison of associating
Property the summation of bio-toxicity that produces when acting solely on body considerably beyond each chemical substance；Summation action
Referring to that multiple Enviromental pollutants simultaneously acts on joint-biotoxicity produced by body is each chemical substance
The summation of the bio-toxicity produced when acting solely on body.
Embodiment 1: detection Cu2+Bio-toxicity
Preparation 10,15,20,25, the Cu of 30mg/L2+Solution, the Cu of detection variable concentrations2+Life
Thing toxicity also calculates its suppression ratio according to formula, obtains concentration-suppression ratio curve.It is calculated Cu2+'s
IC50.The results are shown in Table 1 and Fig. 2.
Embodiment 2: detection Cd2+Bio-toxicity
Preparation 10,15,20,25, the Cd of 30mg/L2+Solution, the Cd of detection variable concentrations2+Life
Thing toxicity also calculates its suppression ratio according to formula, obtains concentration-suppression ratio curve.It is calculated Cd2+'s
IC50.The results are shown in Table 1 and Fig. 3.
Embodiment 3: the bio-toxicity of detection 3,5-chlorophenesic acid (DCP)
Preparation 10,15,20,25, the 3 of 30mg/L (concentration), 5-chlorophenesic acid, according to embodiment 1
Method detection variable concentrations 3, the bio-toxicity of 5-chlorophenesic acid (DCP), and calculate it according to formula
Suppression ratio, obtains concentration-suppression ratio curve.It is calculated the IC50 of DCP.The results are shown in Table 1 and Fig. 4.
Embodiment 4: the bio-toxicity of detection orthene
Preparation 10,20,30,40, the agricultural chemical insecticide orthene of 50mg/L (concentration), according to
The bio-toxicity of orthene of the method detection variable concentrations of embodiment 1, and calculate it according to formula and press down
Rate processed, obtains concentration-suppression ratio curve.It is calculated the IC50 of orthene.The results are shown in Table 1 and scheme
5。
Embodiment 5: the bio-toxicity of detection herbicide
Preparation 10,15,20,30, the herbicide of 40mg/L (concentration), according to embodiment 1
The bio-toxicity of ametryn of method detection variable concentrations, and calculate its suppression ratio according to formula, obtain dense
Degree-suppression ratio curve.It is calculated Cd2+IC50.It is calculated the IC50 of ametryn.The results are shown in Table 1
And Fig. 6.
Table 1: the IC50 value of single poisonous substance
| Poisonous substance | Cu2+ | Cd2+ | 3,5-chlorophenesic acid (DCP) | Orthene | Ametryn |
| IC50(mg/L) | 25.50 | 16.50 | 17.10 | 28.30 | 18.67 |
Embodiment 6: the bio-toxicity of actual waste water is detected
The bio-toxicity of refuse landfill waste water, electroplating wastewater and laboratory organic wastewater is detected,
Detection method is with above example 1, and its suppression ratio is respectively 39.83%, 48.15% and 51.77%, result
See Fig. 7.
Embodiment 7: detection Cu2+And Cd2+Joint-biotoxicity
By Cu identical for toxic unit2+And Cd2+It is mixed and made into Cu by 1:12+-Cd2+Mixing sample, selected
The Concentraton gradient taken is 20%TU, 40%TU, 60%TU, 80%TU, 100%TU, 120%TU,
Detect the suppression ratio of this mixing sample according to the detection method of joint-biotoxicity, and draw concentration-suppression ratio
Curve, it is thus achieved that Cu2+-Cd2+The IC50 of mixing sampleobs, and contrast IC50obsAnd IC50expSize,
Determine Cu2+And Cd2+Joint toxicity type.The results are shown in Table 2.
Embodiment 8: detection Cu2+Joint-biotoxicity with DCP
Cu is prepared according to the method for embodiment 72+-DCP mixing sample, according to the detection of joint-biotoxicity
Method detects the suppression ratio of this mixing sample, and draws concentration-suppression ratio curve, it is thus achieved that Cu2+-DCP mixes
Close the IC50 of sampleobs, and contrast IC50obsAnd IC50expSize, determine Cu2+Connection with DCP
Close pathotype.The results are shown in Table 2.
Embodiment 9: detection orthene and the joint-biotoxicity of DCP
Orthene-DCP mixing sample is prepared, according to joint-biotoxicity according to the method for embodiment 7
Detection method detect the suppression ratio of this mixing sample, and draw concentration-suppression ratio curve, it is thus achieved that acetyl first
The IC50 of amine phosphorus-DCP mixing sampleobs, and contrast IC50obsAnd IC50expSize, determine acetyl
The joint toxicity type of Bayer 71628 and DCP.The results are shown in Table 2.
Embodiment 10: detection orthene and Cu2+Joint-biotoxicity
Orthene-Cu is prepared according to the method for embodiment 72+Mixing sample, according to joint-biotoxicity
Detection method detect the suppression ratio of this mixing sample, and draw concentration-suppression ratio curve, it is thus achieved that acetyl first
Amine phosphorus-Cu2+The IC50 of mixing sampleobs, and contrast IC50obsAnd IC50expSize, determine acetyl
Bayer 71628 and Cu2+Joint toxicity type.The results are shown in Table 2.
Embodiment 11: detection orthene and the joint-biotoxicity of ametryn
Orthene-ametryn mixing sample is prepared, according to the biological poison of associating according to the method for embodiment 7
The detection method of property detects the suppression ratio of this mixing sample, and draws concentration-suppression ratio curve, it is thus achieved that acetyl
The IC50 of Bayer 71628-ametryn mixing sampleobs, and contrast IC50obsAnd IC50expSize, determine
The joint toxicity type of orthene and ametryn.The results are shown in Table 2.
Table 2 mixes toxicity detection and the type of action thereof of poisonous substance
Embodiment 12: single bacterium and the contrast of mixed vaccine biosensor detection bio-toxicity
Utilize single escherichia coli, single bacillus subtilis and the single strain of single saccharomyces cerevisiae preparation raw
Thing sensor, and the Cu of DCP, 10mg/L to 10mg/L respectively2+With the ametryn of 25mg/L
Bio-toxicity detects.By experimental result respectively with embodiment 3, embodiment 1 and the result of embodiment 5
Contrast, the results are shown in Table 3.
The preparation method of single strain bio sensor is basic phase with the preparation method of mixed vaccine biosensor
With, the difference is that only, single strain bio sensor comprises only a kind of thalline, i.e. contain escherichia coli,
One in bacillus subtilis and yeast, and mixed vaccine biosensor contains three kinds of bacterium, i.e. large intestine
Bacillus, bacillus subtilis and saccharomyces cerevisiae.Additionally escherichia coli, bacillus subtilis and yeast
Yeast culture, microorganism collection, thalline dispersion liquid preparation with embodiment 1.Test result such as table 3.By
Following table understands, and the detection sensitivity of single strain bio sensor is either to organic poison DCP, heavy metal
Ion Cu2+Or organic agricultural chemicals ametryn is all not as mixed vaccine biosensor.
Table 3: mixed vaccine
Obviously, the above embodiment of the present invention is only for clearly demonstrating example of the present invention, and
It is not the restriction to embodiments of the present invention, for those of ordinary skill in the field,
Can also make other changes in different forms on the basis of described above, here cannot be to all
Embodiment give exhaustive, every belong to the obvious change that technical scheme extended out
Change or change the row still in protection scope of the present invention.
Claims (8)
1. the method for a mixed vaccine biosensor detection pollutant bio-toxicity, it is characterised in that bag
Include following steps:
1) preparing microbial inoculum, it includes yeast liquid, Escherichia coli bacteria liquid and bacillus subtilis bacterium
Liquid；
2) it is coated on fexible film after described microbial inoculum being mixed homogeneously with mixed sols, prepares micro-
Biofilm；
3) described microbial thin film is assembled on electrode, i.e. prepares mixed vaccine biosensor；
4) detection testing sample；
5) bio-toxicity of sample to be tested is evaluated.
The method of detection pollutant bio-toxicity the most according to claim 1, it is characterised in that system
The method of standby microbial inoculum is:
1) microbial inoculant is cultivated to exponential phase in culture medium；
2) thalline is collected, it is thus achieved that bacterial sediment；
3) washing bacterial sediment；
4) disperse bacterial sediment with dispersion liquid, make microorganism dispersion liquid；
5) it is mixed in proportion different types of microorganism dispersion liquid, makes microbial inoculum.
The method of detection pollutant bio-toxicity the most according to claim 1, it is characterised in that big
Enterobacteria: bacillus subtilis: yeast is 1-3:1-3:1.
The method of detection pollutant bio-toxicity the most according to claim 1, it is characterised in that big
Enterobacteria: bacillus subtilis: yeast is 2:2:1.
Mixed vaccine method for biosensor the most according to claim 1, it is characterised in that mix molten
Glue is polyvinyl alcohol colloidal sol, polyvinyl alcohol-4-ethylpyridine graft polymers colloidal sol, polyvinyl alcohol-sodium sulfate
Colloidal sol or polyvinyl alcohol-alginate mixed sols.
The method of detection pollutant bio-toxicity the most according to claim 1, it is characterised in that institute
Stating fexible film is polyurethane film, polydimethylsiloxanefilm film, polyvinyl chloride film or poly-
Ethylene glycol terephthalate thin film.
The method of detection pollutant bio-toxicity the most according to claim 1, it is characterised in that institute
State sample to be tested detection method to comprise the steps:
1) breathing medium is added；
2) apply voltage, and detect electric current；
3) add benzoquinone solution, and detect electric current, it is thus achieved that the first current intensity；
4) sample to be tested is added；
5) detection electric current, it is thus achieved that the second current intensity.
The method of detection pollutant bio-toxicity the most according to claim 1, it is characterised in that use
Suppression ratio evaluates the bio-toxicity of sample to be tested, and the computing formula of described suppression ratio is as follows:
Suppression ratio (%)=(1-I2/I1) × 100%
Wherein, I1It is the first current intensity, I2It is the second current intensity.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107631485A (en) * | 2017-09-28 | 2018-01-26 | 深圳市鑫汇达机械设计有限公司 | A kind of solar water heater water delivery device |
| CN107703201A (en) * | 2017-09-28 | 2018-02-16 | 韦彩霞 | A kind of aquaculture persticide residue early warning system |
| CN108226260A (en) * | 2017-12-27 | 2018-06-29 | 潘彦伶 | A kind of heavy metal quick-check sensor of cosmetics |
| CN110542749A (en) * | 2019-08-23 | 2019-12-06 | 广州环投环境服务有限公司 | Landfill leachate toxicity detection method |
| CN110923130A (en) * | 2019-10-14 | 2020-03-27 | 嘉兴学院 | Microbial sensor and preparation method and application thereof |
| CN115044473A (en) * | 2022-05-07 | 2022-09-13 | 北京航空航天大学 | Preparation process of electrochemically active bacterium freeze-dried powder and method for detecting water quality biotoxicity in real time |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101261242A (en) * | 2008-04-21 | 2008-09-10 | 同济大学 | Water quality toxicity biosensor microorganism electrode preparation method |
| CN103940883A (en) * | 2013-01-21 | 2014-07-23 | 中国科学院理化技术研究所 | Preparation method, application, device and detection method of one-time microbial film sensor for rapid detection of biotoxicity of water |
-
2015
- 2015-02-04 CN CN201510058337.2A patent/CN105986000B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101261242A (en) * | 2008-04-21 | 2008-09-10 | 同济大学 | Water quality toxicity biosensor microorganism electrode preparation method |
| CN103940883A (en) * | 2013-01-21 | 2014-07-23 | 中国科学院理化技术研究所 | Preparation method, application, device and detection method of one-time microbial film sensor for rapid detection of biotoxicity of water |
Non-Patent Citations (2)
| Title |
|---|
| 丁钰力等: "基于枯草芽孢杆菌微生物传感器的毒性分析", 《中国环境科学》 * |
| 贺莹等: "CellSense生物传感器毒性分析系统应用研究", 《生态毒理学报》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107631485A (en) * | 2017-09-28 | 2018-01-26 | 深圳市鑫汇达机械设计有限公司 | A kind of solar water heater water delivery device |
| CN107703201A (en) * | 2017-09-28 | 2018-02-16 | 韦彩霞 | A kind of aquaculture persticide residue early warning system |
| CN108226260A (en) * | 2017-12-27 | 2018-06-29 | 潘彦伶 | A kind of heavy metal quick-check sensor of cosmetics |
| CN110542749A (en) * | 2019-08-23 | 2019-12-06 | 广州环投环境服务有限公司 | Landfill leachate toxicity detection method |
| CN110923130A (en) * | 2019-10-14 | 2020-03-27 | 嘉兴学院 | Microbial sensor and preparation method and application thereof |
| CN110923130B (en) * | 2019-10-14 | 2023-08-18 | 嘉兴学院 | Microbial sensor and preparation method and application thereof |
| CN115044473A (en) * | 2022-05-07 | 2022-09-13 | 北京航空航天大学 | Preparation process of electrochemically active bacterium freeze-dried powder and method for detecting water quality biotoxicity in real time |
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