CN103043799B - Method for using organic tin-degrading bacteria agent and remediation plants to treat organic tin polluted water - Google Patents
Method for using organic tin-degrading bacteria agent and remediation plants to treat organic tin polluted water Download PDFInfo
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Abstract
The invention discloses a method for using organic tin-degrading bacteria agent and remediation plants to treat organic tin polluted water. According to the invention, a large amount of organic tin-degrading bacteria are obtained through fermentation technology, and the bacteria are dried to obtain active bacteria agent which can be stored for a long time. Under the condition of planting remediation plants, the organic tin-degrading bacteria are added to remove organic tin by using the combined processing technology of organic tin-degrading bacteria agent and remediation plants. The method has the advantages that the combined organic tin pollutant processing technology of organic tin-degrading bacteria agent and remediation plants is adopted, the treatment efficiency is high, the cost is low, the method does not cause secondary pollution, the technical effect of the method is better than single microbial purifying, the quantity of the organic tin-degrading bacteria required to be added into water is small, and the remediation plants also has the feature of landscaping.
Description
Technical field
The invention belongs to the environment bioengineering technical field, particularly a kind of method of utilizing organotin degradation bacterial agent and rehabilitation plant to process the organotin pollution water body.
Background technology
Organotin is used as catalyzer, sterilant, sterilant, weedicide, rust-preventive agent, marine finish, wood preservative, anti-mildew agent for textiles and macromolecular material stablizer etc. in large quantities, one of organo-metallic of most widely used general and quantity discharged maximum, ten thousand tons of annual usage quantity more than tens.The organotin pollution thing has the characteristic of heavy metal and persistence organic pollutant concurrently, bioconcentration with high toxicity, persistence and height, can cause comprising the species generation sex distortion of mollusk, fish and Mammals, and can jeopardize the ecosystem by the enrichment of food chain.In long-term production and use procedure, the organotin pollution thing finally can enter in water body continuously by various blowdown approach, and the organotin water pollution becomes the very severe environmental problem that China faces.
Certain micro-organisms is because organotin being had to resistance or can being used organotin as carbon source, thereby becomes the main force of water body organotin degraded.To the research of this aspect, existing report mainly lays particular emphasis on screening and the influence factor of degradation bacteria and inquires into.Research method is divided three classes: 1. utilize the high-efficiency strain of screening purifying, the organotin in water sample is degraded.Research shows, the metal ion coexisted in temperature, dissolved oxygen concentration, degradation bacteria kind, degraded system and small molecular organic acid etc. all can exert an influence to the microbiological deterioration effect of organotin.2. utilize immobilized reagent that degradation bacteria is prepared as to bead aftertreatment organotin.The advantage of the method is to reduce the loss of degradation bacteria, and solid-liquid separation is easy, still, is adsorbed in hard degradation of organotin in immobilized reagent.The organotin of 3. take in settling or municipal sludge is to process object, analyzes indigenous microorganism in settling or municipal sludge and the inoculating microbe degradation effect to organotin.But the degradation bacteria performance of researchist's seed selection both at home and abroad is all not ideal enough at present; And, when single degradation bacteria is processed the water body organotin, exist easily run off, degrading enzyme is single, to shortcomings such as the organotin toxicity adaptive phase are partially long.
Summary of the invention
The shortcoming that the object of the invention is to overcome prior art, with not enough, provides a kind of method of utilizing organotin degradation bacterial agent and rehabilitation plant to process the organotin pollution water body.
Purpose of the present invention is achieved through the following technical solutions: a kind of method of utilizing organotin degradation bacterial agent and rehabilitation plant to process the organotin pollution water body comprises the steps:
(1) organotin degradation bacteria liquid is inoculated in substratum by 1～10% volume ratio, is placed in fermentor tank and cultivates, when the nectar degree of organotin degradation bacteria reaches 5～12g/L, the results thalline, drying, obtain the organotin degradation bacterium preparation;
(2) according to processing water body organotin content, ratio in every strain plant treatment 1～20mg organotin, the plantation rehabilitation plant, each season adds by 5～70mg/L the organotin degradation bacterium preparation that step (1) makes, accelerate the organotin degradation bacterium preparation and grow in plant rhizosphere or bed mud, and realize the organotin degradation bacteria with rehabilitation plant to the quick-acting reparations of combining of organotin;
In step (1):
Described organotin degradation bacteria liquid adopts following methods to be prepared: in organotin degradation bacteria inclined-plane, picking organotin degradation bacteria is inoculated in seed culture medium by 1～50mg/L, shaking culture 10～24h in 50～200r/min shaking table, obtain organotin degradation bacteria liquid;
Described organotin degradation bacteria is preferably bacillus thuringiensis (Bacillus thuringiensis);
The composition of described seed culture medium is extractum carnis 3～10g/L, peptone 2～10g/L and NaCl 0～8g/L;
Described medium optimization is beef-protein medium;
The composition of described beef-protein medium is extractum carnis 3～12g/L, peptone 3～10g/L and NaCl 0～8g/L;
Described 1～10% volume ratio refers to 1～10% of volume that the volume of organotin degradation bacteria liquid is substratum;
Described cultivation is preferably carried out in dissolved oxygen per-cent is 30 ℃ of 80wt%, pH 5.8, temperature;
Described results thalline is preferably undertaken by mode centrifugal or that filter;
Described centrifugal speed is preferably 3000～8000rpm;
Described drying is preferably 40～100 ℃ of dryings or vacuum lyophilization;
Rehabilitation plant described in step (2) is preferably rib water shield (Pontederia cordata);
Preferably, step adds sucrose fatty ester, rhamnolipid or supplemental oxygen in (2), accelerates the reparation progress;
It is 1～4mg/L that described supplemental oxygen preferably maintains dissolved oxygen concentration;
The dosage of described sucrose fatty ester or rhamnolipid is preferably every liter of processing water body or the per kilogram water body deposit adds 2～60mg sucrose fatty ester or rhamnolipid;
Invention mechanism of the present invention: plant to the bio-transformation of high molecular organotin and transportcapacity a little less than, when organotin degradation bacteria and rehabilitation plant combined utilization, can realize both mutual supplement with each other's advantages.Plant rhizosphere is given birth to site for the organotin degradation bacteria is providing, and has improved the biomass of repairing place organotin degradation bacteria; And coexisting of organotin degradation bacteria and rehabilitation plant is conducive to accelerate the removing toxic substances of high toxicity organotin and thoroughly degraded.Rehabilitation plant and organotin degradation bacteria all can be carried out biological adsorption, cylinder accumulation and progressively de-alkyl degraded to organotin.Degradation pathway is as follows: R
4snX → R
3snX → R
2snX
2→ RSnX
3→ Sn, wherein R is the alkyl such as methyl, ethyl, butyl, cyclohexyl, phenyl or octyl group; Sn is inorganic tin; X is the groups such as inorganic acid radical, organic acid, oxygen or haloid element.
The molecular weight of organotin is larger, usually is solid-state or oily is liquid, water-soluble low, this characteristic limitations of organotin contacting of organotin and rehabilitation plant and organotin degradation bacteria, be unfavorable for the biological degradation of organotin.Tensio-active agent can make organotin dispersion, solubilising, emulsification as sucrose fatty ester and rhamnolipid, and then improves wetting ability and the bioavailability of organotin, be conducive to rehabilitation plant and organotin degradation bacteria to its absorption with utilize.
The present invention has following advantage and effect with respect to prior art:
(1) the present invention obtains a large amount of organotin degradation bacteria by fermentation technique, but and thalline is carried out to the dry rear active microbial inoculum that obtains long storage time.In the situation that the plantation rehabilitation plant adds the organotin degradation bacteria, utilize organotin degradation bacterial agent and rehabilitation plant combination treatment technology, realize the removal of organotin.Plant to the bio-transformation of high molecular organotin and transportcapacity a little less than, when organotin degradation bacteria and rehabilitation plant combined utilization, can realize both mutual supplement with each other's advantages; Plant rhizosphere is given birth to site for the organotin degradation bacteria is providing, and has improved the biomass of repairing place organotin degradation bacteria; And coexisting of organotin degradation bacteria and rehabilitation plant is conducive to accelerate the removing toxic substances of high toxicity organotin and thoroughly degraded.By adding tensio-active agent, can effectively promote dispersion and the emulsification of organotin, thereby acceleration rehabilitation plant and organotin degradation bacteria absorb and degraded to it.The present invention combines the advantage of rehabilitation plant, organotin degradation bacteria and tensio-active agent effectively, and the water body organotin is carried out to the quick bio processing.
(2) the present invention utilizes the technology of organotin degradation bacterial agent and rehabilitation plant combined purifying organotin pollution thing more superior than independent microbially decontaminate technique effect, still less, rehabilitation plant also has the feature of the landscape of beautifying to the required bacterium amount of rendering to the organotin degradation bacteria in water.
(3) the present invention utilizes the organotin degradation bacterial agent to compare with heat treating process with chemical method with the technology of rehabilitation plant combined purifying organotin pollution thing, has that processing efficiency is high, cost is low and the advantage such as non-secondary pollution.
The accompanying drawing explanation
Fig. 1 is the interpretation of result figure of different dry drying method on the impact of organotin degradation bacterium preparation activity in comparative example 1, wherein: 1-contrast, 2-vacuum lyophilization, 3-air blast heated drying (40 ℃), 4-air blast heated drying (60 ℃), 5-air blast heated drying (100 ℃), 6-Vacuum Heat drying.
Fig. 2 is the figure as a result that in embodiment 1, the different treatment method is processed organotin, wherein :-◆-for adopting the figure as a result of genus bacillus-rib water shield combination treatment organotin,-■-for adopting genus bacillus, process the figure as a result of organotin ,-△-for adopting the rib water shield, process the figure as a result of organotin.
Fig. 3 adopts the gas chromatogram of the degraded product of organotin degradation bacteria-rehabilitation plant combination treatment organotin in embodiment 1.
Fig. 4 is the as a result figure of different treatment method to the processing of organotin pollution harbour sewage in embodiment 2, wherein: a is for adopting the figure as a result of genus bacillus-rib water shield combination treatment organotin pollution harbour sewage, b is for adopting genus bacillus to process the figure as a result of organotin pollution harbour sewage, and c is for adopting the rib water shield to process the figure as a result of organotin pollution harbour sewage.
Fig. 5 is the figure as a result that in embodiment 3, the different treatment method is processed coastal waters bed mud organotin, wherein :-◆-for adopting the figure as a result of genus bacillus-rib water shield combination treatment coastal waters bed mud organotin,-■-for adopting genus bacillus, process the figure as a result of coastal waters bed mud organotin ,-△-for adopting the rib water shield, process the figure as a result of coastal waters bed mud organotin.
Fig. 6 is that in comparative example 2, sucrose fatty ester gushes the figure as a result of the promoter action of settling organotin biological restoration to river, wherein :--for not adding sucrose fatty ester, river is not gushed to the results of comparison figure of settling organotin biological restoration, the figure as a result of the promoter action of settling organotin biological restoration is gushed for the 2mg/kg sucrose fatty ester in-*-to river,-△-for the 50mg/kg sucrose fatty ester, river is gushed to the figure as a result of the promoter action of settling organotin biological restoration,-◆-river is gushed to the figure as a result of the promoter action of settling organotin biological restoration for the 60mg/kg sucrose fatty ester.
Fig. 7 is usingd the structural representation that the PVC plate gushes as material construction 5 lattice deflector types simulations rivers in embodiment 1.
Embodiment
Below in conjunction with embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are not limited to this.
Embodiment 1: organotin degradation bacteria and the rehabilitation plant combination treatment to organotin pollution river water burst body
(1) in organotin degradation bacteria inclined-plane picking organotin degradation bacteria bacillus thuringiensis (Bacillusthuringiensis practices the river settling from the Gui Yuzhen of Guangdong Province.Lower same.) be inoculated into seed culture medium (seed culture based component: extractum carnis 3g/L, peptone 10g/L, NaCl 5g/L by 25mg/L.) in, after shaking culture 10h, obtain organotin degradation bacteria liquid in the 50r/min shaking table; Get organotin degradation bacteria liquid and be inoculated in respectively by table 1 to become in the substratum of assignment system by 1% volume ratio, be placed in fermentor tank and cultivate, investigate the effect of medium component to yeast culture.When the nectar degree of organotin degradation bacteria reaches 5g/L, in the centrifugal results thalline of 3000rpm, 40 ℃ of dryings, obtain gemma bacillus agent;
Table 1 fermentation culture based component
Described cultivation is carried out in dissolved oxygen per-cent is 30 ℃ of 80wt%, pH 5.8, temperature;
(2) according to processing water body organotin content, ratio in every strain plant treatment 1mg organotin, the plantation rehabilitation plant, each season adds by 5mg/L the gemma bacillus agent that step (1) makes, accelerate gemma bacillus agent and grow in plant rhizosphere or bed mud, and realize the organotin degradation bacteria with rehabilitation plant to the quick-acting reparations of combining of organotin;
Rehabilitation plant: rib water shield (Pontederia cordata), screen acquisition by the inventor: take the Yong Ji greenery patches, river of Guangzhou and surrounding cities buys plant as the herborization of plant sampling point and in flowers market, according to roots of plants length, increment, resistant effect and the removal effect of organotin is screened to rehabilitation plant; Select roots of plants to grow, increment is large, resistant is effective and the good rehabilitation plant to the removal effect of organotin;
Water sampling: be collected in Guangzhou Che Po and gush, be transported to behind laboratory and analyze water quality (its pH, BOD at once
5, COD
cr, TN and TP analytical results be respectively 6.9,28mg/L, 49mg/L, 13.2mg/L and 1.6mg/L), and carry out the organotin degradation bacteria of water sample-plant Combined Processing experiment;
The rib water shield is cultivated: the rib water shield is fixed in the Glass Containers that volume is 20L, adds the 14L plant nutrition liquid and is cultivated, and every 3d changes liquid once, cultivates altogether 14d, chooses the degradation experiment that good, the of the same size rib water shield of growth conditions carries out organotin.Indoor day temperature of plant-growth is 28 ± 2 ℃, and night, temperature was 24 ± 2 ℃, and light application time is 14h/d, and intensity of illumination is 5000～6500lx;
Organotin degradation bacteria and the rehabilitation plant combination treatment to organotin pollution river water burst sample: the PVC plate of usining gushes (as shown in Figure 7) as material construction 5 lattice deflector type simulation rivers, and the specification that every lattice are gushed in river is 200cm * 15cm * 30cm.Get in river water burst sample is positioned over simulation river gushes, to available depth be 20cm; Be taken at the rib water shield of cultivating 14d in nutritive medium, with tap water, rinse rib water shield root system well, then adopt cystose that the rib water shield is fixed in water, planting plants 5 strains of every lattice river course, totally 25 strains; Add organotin in intending the water sample of processing, the concentration that makes organotin is 200 μ g/L, and adds genus bacillus, and the concentration that makes genus bacillus in water sample is 50mg/L; Gush end in the 5th lattice simulation river and place water pump, water sample pump to the 1 lattice river course, make water sample gush middle formation circulation in the simulation river; The concentration of the remaining organotin of every 2d sampling analysis (utilizes high performance liquid chromatograph to carry out the TPhT detection, testing conditions: C
18reversed-phase column (150mm), UV-detector (detecting wavelength 206nm); Moving phase V
methyl alcohol: V
water: V
0.1% trifluoroacetic acid solution=50:45:5, flow velocity is 1mL/min; Be 10min analysis time; Sample size 20 μ L.), measure altogether 14d; Separately establish the control experiment of organotin degradation bacteria, rehabilitation plant individual curing organotin, and do not have planting plants not add the blank of organotin degradation bacteria;
The organotin biodegradable product is analyzed: the water sample 10mL that gets organotin degradation bacteria and rehabilitation plant combination treatment 14d, add the ultrasonic extraction of 10mL normal hexane at every turn, coextraction 2 times, each 30min, the organic phase that merges 2 extractions, rotate evaporate to dryness under 35 ℃, use the 6mL methanol-eluted fractions, add the 4mL NaAc_HAc buffer solution, the NaDDTC solution of 1mL 2%, stratification after the ultrasonic 15min of 5mL normal hexane, get upper organic phase and carry out the GC-MS analysis (condition that GC-MS analyzes: adopt Rxi-5MS GC post (30m * 0.25mm * 0.25 μ m) to carry out product analysis in the GC-MS instrument, carrier gas is high-purity helium, flow velocity is that the 1.1mL/min. column temperature maintains 1.5min in 50 ℃, be warming up to 300 ℃ and to maintain the 4min. interface temperature be 280 ℃ by the speed of 10 ℃/min subsequently, ion source temperature is 250 ℃, Selective ion monitoring (SIM) mode, sample size is 2 μ L),
Organotin degradation bacteria quantity is with active: get rehabilitation plant rhizosphere water sample, in sterilized water, dilute respectively 10
4, 10
5, 10
6doubly, get bacterium liquid 0.2mL and be inoculated in the extractum carnis culture plate, cultivate 24h in 37 ℃ of incubators after, calculate colony number, thereby measure the quantity of genus bacillus and rib water shield combination treatment organotin process viable bacteria body; Adopt BIOLOG technical measurement organotin degradation bacteria activity;
Result shows, the culture effect ideal of the substratum of each concentration range that experiment is selected to genus bacillus, and after cultivating, cell concentration is all over 5g/L; Genus bacillus and rib water shield combination treatment technique effect ideal, be better than the independent role of two species; After processing 14d, the clearance of organotin is up to 73%(Fig. 2), triphenyltin is degraded to tin diphenyl, a phenyltin (Fig. 3) and inorganic tin effectively.
Embodiment 2: organotin degradation bacteria and the rehabilitation plant combination treatment to organotin pollution harbour sewage
(1) in organotin degradation bacteria inclined-plane, picking organotin degradation bacteria bacillus thuringiensis (Bacillusthuringiensis) is inoculated into seed culture medium (seed culture based component: extractum carnis 10g/L, peptone 2g/L, NaCl 0g/L by 1mg/L.) in, after shaking culture 15h, obtain organotin degradation bacteria liquid in the 100r/min shaking table; Get organotin degradation bacteria liquid and be inoculated in substratum (medium component: extractum carnis 12g/L, peptone 3g/L, NaCl 0g/L by 5% volume ratio.) in, be placed in fermentor tank and cultivate, when the nectar degree of organotin degradation bacteria reaches 9g/L, centrifugal results thalline under the condition of 3000rpm, 100 ℃ of dryings, obtain gemma bacillus agent;
Described cultivation is carried out in dissolved oxygen per-cent is 30 ℃ of 80wt%, pH 5.8, temperature;
(2) according to processing water body organotin content, ratio in every strain plant treatment 10mg organotin, the plantation rehabilitation plant, each season adds by 30mg/L the gemma bacillus agent that step (1) makes, accelerate gemma bacillus agent and grow in plant rhizosphere or bed mud, and realize the organotin degradation bacteria with rehabilitation plant to the quick-acting reparations of combining of organotin;
Rehabilitation plant: rib water shield (Pontederia cordata), screen acquisition by the inventor: take the Yong Ji greenery patches, river of Guangzhou and surrounding cities buys plant as the herborization of plant sampling point and in flowers market, according to roots of plants length, increment, resistant effect and the removal effect of organotin is screened to rehabilitation plant; Select roots of plants to grow, increment is large, resistant is effective and the good rehabilitation plant to the removal effect of organotin;
The specification that the PVC plate of take gushes every lattice as material construction 5 lattice deflector type simulation Yong， rivers, river is 200cm * 15cm * 30cm.Get in river water burst sample is positioned over simulation river gushes, to available depth be 20cm.Be taken at the rib water shield of cultivating 14d in nutritive medium, rinse rib water shield root system well with tap water, then utilize cystose that the rib water shield is fixed in and gathers in the water sample of port, Huangpu, Guangzhou harbour (starting point concentration of organotin is 10.41 μ g/L), planting plants 5 strains of every lattice river course, totally 25 strains, add genus bacillus, the concentration that makes genus bacillus in water sample is 50mg/L.Gush end in the 5th lattice simulation river and place water pump, water sample pump to the 1 lattice river course, make water sample gush middle formation circulation in the simulation river, in the concentration of the remaining organotin of 14d sampling analysis.Separately establish the control experiment of organotin degradation bacteria, rehabilitation plant individual curing organotin, and do not have planting plants not add the blank of organotin degradation bacteria;
Result shows, genus bacillus and rib water shield combination treatment technique effect ideal are better than the independent role of two species; After processing 14d, the clearance of organotin is up to 97%(Fig. 4).
Embodiment 3: organotin degradation bacteria and the rehabilitation plant combination treatment to organotin pollution coastal waters bed mud
(1) in organotin degradation bacteria inclined-plane, picking organotin degradation bacteria bacillus thuringiensis (Bacillusthuringiensis) is inoculated into seed culture medium (seed culture based component: extractum carnis 6g/L, peptone 5g/L, NaCl 8g/L by 50mg/L.) in, after shaking culture 24h, obtain organotin degradation bacteria liquid in the 200r/min shaking table; Get organotin degradation bacteria liquid and be inoculated in substratum (medium component: extractum carnis 8g/L, peptone 8g/L, NaCl 8g/L by 10% volume ratio.) in, be placed in fermentor tank and cultivate, when the nectar degree of organotin degradation bacteria reaches 12g/L, centrifugal results thalline under the condition of 3000rpm, 60 ℃ of dryings, obtain gemma bacillus agent;
Described cultivation is carried out in dissolved oxygen per-cent is 30 ℃ of 80wt%, pH 5.8, temperature;
(2) according to processing water body organotin content, ratio in every strain plant treatment 20mg organotin, the plantation rehabilitation plant, each season adds by 70mg/L the gemma bacillus agent that step (1) makes, accelerate gemma bacillus agent and grow in plant rhizosphere or bed mud, and realize the organotin degradation bacteria with rehabilitation plant to the quick-acting reparations of combining of organotin;
Rehabilitation plant: rib water shield (Pontederia cordata), screen acquisition by the inventor: take the Yong Ji greenery patches, river of Guangzhou and surrounding cities buys plant as the herborization of plant sampling point and in flowers market, according to roots of plants length, increment, resistant effect and the removal effect of organotin is screened to rehabilitation plant; Select roots of plants to grow, increment is large, resistant is effective and the good rehabilitation plant to the removal effect of organotin;
The coastal waters bed mud gathers: experiment was collected in Guangzhou Huang Pugang the same day, was transported to laboratory post analysis bed mud organotin content (1.87mg/kg), and carried out the organotin degradation bacteria of bed mud organotin-plant Combined Processing experiment;
Organotin degradation bacteria and the rehabilitation plant combination treatment to coastal waters bed mud organotin: get bed mud 10kg in Glass Containers, cover distilled water 5L on matrix, add genus bacillus 1g and mix all, rib water shield 5 strains that the plantation plant height is 70cm.Every 2d gets the concentration that rib water shield rhizosphere bed mud is analyzed remaining organotin, measures altogether 14d.Separately establish the contrast of organotin degradation bacteria, rehabilitation plant individual curing organotin, and do not have planting plants not add the blank of organotin degradation bacteria;
Found that, genus bacillus and rib water shield not only can effectively be removed the organotin in water, and the result of Fig. 5 also shows, this organotin degradation bacteria-rehabilitation plant combination treatment technology can also be processed the organotin in the bed mud of coastal waters effectively; When 14d, the clearance of bed mud organotin is reached to 67%, each sampling time point all is better than the effect of genus bacillus or rib water shield individual curing.
The comparative example 1: the preparation of organotin degradation bacterium preparation and to the degraded of organotin
1 material
Organotin degradation bacteria: bacillus thuringiensis (Bacillus thuringiensis);
Seed culture medium: extractum carnis 5g/L, peptone 8g/L, NaCl 5g/L;
Substratum: extractum carnis 7g/L, peptone 5g/L, NaCl 3g/L;
2 methods
Fermentation process: in organotin degradation bacteria inclined-plane, picking organotin degradation bacteria is inoculated in seed culture medium by 20mg/L, respectively at 50,100 and the 200r/min shaking table in shaking culture 10,12 and 24h, obtain organotin degradation bacteria liquid; Get organotin degradation bacteria liquid and be inoculated in substratum by 1%, 5% and 10% volume ratio, be placed in fermentor tank and cultivate, the dissolved oxygen per-cent in the controlled fermentation tank is 80%, pH 5.8, and temperature is 30 ℃; When the nectar degree of organotin degradation bacteria reaches 8g/L, centrifugal results thalline under the condition of 3000rpm, adopt respectively the methods such as lyophilize, air blast heated drying and Vacuum Heat drying to obtain the organotin degradation bacterium preparation;
Vacuum lyophilization: thalline is carried out to vacuum lyophilization in-55 ℃;
Air blast heated drying: under the condition of air blast, in 40 ℃, 60 ℃, 100 ℃ dry thalline;
Vacuum Heat drying: under the condition that is 0.08MPa in vacuum tightness, in 60 ℃ of dry thalline;
Drying conditions determination methods: get and adopt the dried microbial inoculum 0.1g of each drying means, dilute suitable multiple in sterilized water after, getting bacterium liquid 0.2mL is inoculated in the extractum carnis culture plate, cultivate 24h in 37 ℃ of incubators after, calculate colony number, thereby measure the quantity of viable bacteria body after each drying means drying; Separately get in the organotin water sample that the dried microbial inoculum of each drying means is added to 200 μ g/L, the ultimate density that makes microbial inoculum is 50mg/L, 5d degrades under 100r/min, 30 ℃ of conditions, and remaining organotin after utilizing the ethyl acetate extraction to degrade, adopt the concentration of the remaining organotin of Syrups by HPLC, there is no dry microbial inoculum, do not carry out control experiment;
The organotin detection method: separator column is C
18reversed-phase column (150mm); Use UV-detector, detect wavelength 206nm; Moving phase V
methyl alcohol: V
water: V
0.1% trifluoroacetic acid solution=50:45:5, flow velocity is 1mL/min; Be 10min analysis time; Sample size 20 μ L;
3 results
Fig. 1 is after bacillus thuringiensis is inoculated in seed culture medium, in the 100r/min shaking table after shaking culture 24h, get seed culture fluid and be inoculated in fermentor tank by 5% volume ratio, the experimental result that after cultivating, the thalline of results carries out the organotin degraded after by various drying meanss.Result shows organotin degradation bacterium preparation superior performance prepared by present technique, under the condition of oscillation treatment, can remove 83% organotin in 5d, and the half sorrow phase that reaches the several years with this pollutant is compared, and degradation speed has accelerated hundred times.The different drying means of microbial inoculum can produce certain impact to the microbial inoculum quantity of viable bacteria body in the effect of organotin and microbial inoculum of degrading, wherein with the best results of vacuum freeze-drying method; But, the heated drying method still can make microbial inoculum to the removal of organotin up to more than 57%; Experimental result and Fig. 1 of other levels of the shaking speed of setting in experiment, seed liquor inoculation volume ratio, air blast heated drying distinguish not obvious.Therefore, aborning, the heated drying method is also feasible.
The comparative example 2: tensio-active agent gushes the promoter action of settling organotin biological restoration to river
1 material
Organotin degradation bacteria: bacillus thuringiensis (Bacillus thuringiensis);
Rehabilitation plant: rib water shield (Pontederia cordata);
Tensio-active agent: sucrose fatty ester;
2 methods
River is gushed settling and is collected in the Dongguan City canal; Get settling 10kg in Glass Containers, cover distilled water 5L on matrix, add genus bacillus 1g, and by 2,50 and the amount of 60mg/kg (settling) add sucrose fatty ester, mix and all plant afterwards rib water shield 5 strains that plant height is 70cm; Every 2d gets the concentration that rib water shield rhizosphere bed mud is analyzed remaining organotin, measures altogether 14d; Separately establish the control experiment that does not add sucrose fatty ester;
3 results
Fig. 6 demonstration, the combined action of genus bacillus and rib water shield, can effectively remove river and gush the organotin in settling; Sucrose fatty ester has and promotes to do this combined action; After adding sucrose fatty ester, the clearance of each sampling time point organotin is approximately high by 10% than the control experiment of not throwing tensio-active agent.Above-described embodiment is preferably embodiment of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under spirit of the present invention and principle, substitutes, combination, simplify; all should be equivalent substitute mode, within being included in protection scope of the present invention.
Claims (6)
1. a method of utilizing organotin degradation bacterial agent and rehabilitation plant to process the organotin pollution water body, is characterized in that comprising the steps:
(1) organotin degradation bacteria liquid is inoculated in substratum by 1～10% volume ratio, is placed in fermentor tank and cultivates, when the nectar degree of organotin degradation bacteria reaches 5～12 g/L, the results thalline, drying, obtain the organotin degradation bacterium preparation;
(2) according to processing water body organotin content, ratio in every strain plant treatment 1～20mg organotin, the plantation rehabilitation plant, each season adds by 5～70 mg/L the organotin degradation bacterium preparation that step (1) makes, accelerate the organotin degradation bacterium preparation and grow in plant rhizosphere or bed mud, and realize the organotin degradation bacteria with rehabilitation plant to the quick-acting reparations of combining of organotin;
Substratum described in step (1) is beef-protein medium, described organotin degradation bacteria is bacillus thuringiensis, described cultivation is carried out in dissolved oxygen per-cent is 30 ℃ of 80 wt %, pH 5.8, temperature, rehabilitation plant described in step (2) is the rib water shield, and adds sucrose fatty ester, rhamnolipid or supplemental oxygen.
2. the method for utilizing organotin degradation bacterial agent and rehabilitation plant to process the organotin pollution water body according to claim 1, it is characterized in that: the composition of described beef-protein medium is extractum carnis 3～12 g/L+ peptone 3～10 g/L+NaCl 0～8 g/L.
3. the method for utilizing organotin degradation bacterial agent and rehabilitation plant to process the organotin pollution water body according to claim 1, it is characterized in that: the organotin degradation bacteria liquid described in step (1) adopts following methods to be prepared: in organotin degradation bacteria inclined-plane, picking organotin degradation bacteria is inoculated in seed culture medium by 1～50 mg/L, shaking culture 10～24 h in 50～200r/min shaking table, obtain organotin degradation bacteria liquid; The composition of described seed culture medium is extractum carnis 3～10 g/L, peptone 2～10 g/L, NaCl 0～8 g/L.
4. the method for utilizing organotin degradation bacterial agent and rehabilitation plant to process the organotin pollution water body according to claim 1, it is characterized in that: the results thalline described in step (1) is undertaken by mode centrifugal or that filter.
5. the method for utilizing organotin degradation bacterial agent and rehabilitation plant to process the organotin pollution water body according to claim 1, it is characterized in that: the drying described in step (1) is 40～100 ℃ of dryings or vacuum lyophilization.
6. the method for utilizing organotin degradation bacterial agent and rehabilitation plant to process the organotin pollution water body according to claim 1, it is characterized in that: it is 1～4mg/L that described supplemental oxygen maintains dissolved oxygen concentration; The dosage of described sucrose fatty ester or rhamnolipid is that every liter of processing water body or per kilogram water body deposit add 2～60 mg sucrose fatty ester or rhamnolipids.
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