CN102745820B - Method for removing MIB and Geosmin in water by using inoculation biofilter - Google Patents
Method for removing MIB and Geosmin in water by using inoculation biofilter Download PDFInfo
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- CN102745820B CN102745820B CN 201210048588 CN201210048588A CN102745820B CN 102745820 B CN102745820 B CN 102745820B CN 201210048588 CN201210048588 CN 201210048588 CN 201210048588 A CN201210048588 A CN 201210048588A CN 102745820 B CN102745820 B CN 102745820B
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Abstract
The invention relates to a method for removing MIB and Geosmin in water by using an inoculation biofilter. The method comprises: preparing an inorganic salt culture media of MIB and Geosmin and a PBS buffer, and adding active carbon to the PBS buffer to obtain a PBS buffer of a biofilm; adopting a pipette to take 10 mL of the biofilm-containing PBS buffer to dissolve in the inorganic salt culture media of the MIB and the Geosmin to carry out selective culture enrichment; adopting a plate streaking method to separate, purify and degrade bacteria to obtain MIB degrading bacteria and Geosmin degrading bacteria; filling cleaned sand in a filtration column, and loading a bacteria liquid containing the MIB degrading bacteria and the Geosmin degrading bacteria on the filtration column, wherein concentration of the bacteria liquid is 1*10<3> cells per mL, an empty bed residence time is 6-60 minutes, and the bacterial liquid flows through the filtration column and circulates for four days, such that the MIB degrading bacteria and the Geosmin degrading bacteria are attached to the surface of the filtration material so as to remove the MIB and the Geosmin in the water. The method of the present invention has the following advantages that: the method is simple; use amounts of ozone and active carbon in the follow-up process are reduced; and characteristics of low cost, good effect, safety and economy are provided.
Description
Technical field
The invention belongs to water-treatment technology field, relate in particular to a kind of biological filter of microbe inoculation bacterial classification that utilizes and remove 2-methyl isoborneol (2-methylisoborneol in tap water, be called for short: MIB) with native smelly element (trans-1,10-dimethyltrans-9-decalol, be called for short: method Geosmin).
Background technology
In recent years, because industrial and agricultural wastewater and sanitary sewage discharge in a large number, the water body the content of nitrogen and phosphorous such as lake, reservoir increase, algae and mushroom amount reproduction, and water body produces smells flavor.At present, tap water is smelt the flavor problem has become the one of the main reasons that causes the human consumer to complain.
Under the impact of mankind's activity, pondage greatly reduces, and water body self-purification ability reduces, the impact of reservoir surrounding environment in addition, and source water water quality runs down.Every the end of spring and the beginning of summer, junior two season autumn late summer, algal grown is vigorous, and source quality obviously descends, and smells flavor serious.The 9th water factory is Beijing topmost water purification supply enterprise, and daily handling ability reaches 1,500,000 tons, and service discharge accounts for 60% of areas of Beijing municipal water supply amount, and the quality of its effluent quality can directly affect the water supply security of Beijing.Annual algae vigorous period, the MIB of this plant effluent and Geosmin content are higher, cause the public's complaint.
Cause the Typical Representative of smelling material---2-methyl isoborneol (MIB, C
11h
20o) and native smelly element (Geosmin, C
12h
22o) being saturated rings tertiary alcohols material, having camphor/native musty, is some blue-green algae and actinomycetic secondary metabolite, olfact extremely low (10ng/L), and traditional water technology (coagulation, filtration and sterilization) is difficult to its removal.
Because the Henry'S coefficient of MIB and Geosmin is low, the cost of stripping technical finesse is higher; Gac has good adsorptivity to MIB and Geosmin, still due to the existence of natural organic matter, larger to the consumption of gac; Due to adding of other material, deep oxidation technology (as carbonic acid gas photochemical catalysis and ultraviolet/hydrogen peroxide treatment technology) can be removed causing in tap water and smell material, but can produce disinfection byproduct (DBP).In the traditional biological filter tank, the specific aim of microorganism is poor, can not remove preferably MIB and Geosmin in water, and improved biological filter can obtain treatment effect preferably.
Summary of the invention
In order to address the above problem, the purpose of this invention is to provide a kind of method simple, reduce ozone and activated carbon dosage in subsequent technique, there is the method that MIB and Geosmin in water are removed in inoculation biological filter that low, the effective advantage of cost has safety, economic characteristics
.
Technical scheme of the present invention is: the method for MIB and Geosmin in inoculation biological filter removal water
, specifically comprise the following steps:
step 1:
1) preparation minimal medium: by the NH of 0.8-1.2g
4nO
3, 0.8-1.2g KH
2pO
4, 0.4-0.6g MgSO
47H
2the KCl of O and 0.2g is dissolved in the distilled water of 1000mL, regulate the pH value to 7.2-7.4, in temperature, be sterilizing 30min under 121 ° of C, with water system needle type filtration device, the MIB that is 20 μ g/L by the concentration of 1-100 μ L under gnotobasis and Geosmin standardized solution add respectively in aseptic 200mL minimal medium, obtain concentration and be respectively 100ng/L, 200ng/L, 500ng/L, 1 μ g/L, 2 μ g/L, 5 μ g/L and the MIB of 10 μ g/L and the minimal medium of Geosmin, standby;
2) preparation LB solid medium: the NaCl of the yeast extract of the peptone of 5-10g, 3-5g and 8-10g is dissolved in the distilled water of 1000mL, the NaOH that is 1mol/L by concentration regulates the pH value to 7.2-7.4, then add the 10-15g agar powder, heating is dissolved agar powder, then cool to room temperature, standby;
3) preparation damping fluid: by Na
2hPO
412H
2o is dissolved in preparation in distilled water and obtains the Na that concentration is 0.2 mol/L
2hPO
4, by NaH
2pO
42H
2o is dissolved in distilled water configuration, and to obtain concentration be 0.2 mol/L NaH
2pO
4; To prepare NaH
2pO
4, Na
2hPO
4after mixing according to the volume ratio of 19:81:200 with water, in temperature, be sterilizing 30min under 121 ° of C, the PBS damping fluid that to obtain concentration be 0.1mol/L, pH=7.4, be stored in dark cold place, standby;
Step 2:
1) get the PBS damping fluid that the 30mL above-mentioned steps prepares, the gac that adds biofilm development maturation in a certain amount of activated carbon filter, concuss 10 min, standing for some time, sterilizing, obtain biomembranous PBS damping fluid, draw and to carry out selectivity in 10mL is dissolved in MIB that the concentration of 200mL is 100ng/L and Geosmin minimal medium containing biomembranous PBS damping fluid and cultivate enrichment with suction pipe, being placed in constant-temperature shaking incubator is 30 ° of C in temperature, under the condition that rotating speed is 160rpm, cultivate, after the substratum muddiness, progressively be transferred to 200ng/L according to 10% inoculum size respectively, 500ng/L, 1 μ g/L, 2 μ g/L, 5 μ g/L, in the MIB of 10 μ g/L and the minimal medium of Geosmin, after each liquid culture stage finishes, draw with the aseptic technique method water sample that 1mL fully mixes, injection fills the 9mL test tube of sterile saline, be mixed into the 1:10 diluent, the diluent 1mL that draws 1:10 annotates the people and fills in the test tube of 9mL sterile saline, is mixed into the 1:100 diluent, is diluted to successively by the same method 1:10
3, 1:10
4, 1:10
5, 1:10
6, 1:10
7diluent, standby, get different dilution diluent 0.2mL with the sterilizing suction pipe, inject respectively in the plate contain freezing solid substratum, with the glass scraper by bacteria suspension in the media surface coating evenly, before use, the glass scraper need be through aseptically process, with after calcination sterilizing on spirit lamp immediately, place cooling, be used alternatingly, do a parallel inoculation during each check, separately with a plate, only pour into the LB solid medium as blank simultaneously, after bacterium liquid absorbs, the upset plate, make bottom surface upwards, be placed in ° C incubator of 30 ° of C ± 1 and cultivate 48h, adopt plate streak separation and purification degraded bacterial classification, obtaining micrococci, Flavobacterium, tyrothricin and pseudomonas is the MIB degradation bacteria, and golden Zymomonas mobilis, Sinorhizobium and Stenotrophomonas are the Geosmin degradation bacteria,
Step 3:
It is highly the Filter column of 100-550mm that sand grains after cleaning is inserted, and the concentration that will contain MIB degradation bacteria and Geosmin degradation bacteria is 1 * 10
3the bacterium liquid of individual/mL, at empty bed residence time 6-60min, flow through Filter column and circulate 4 days, makes MIB degradation bacteria and Geosmin degradation bacteria be attached to described filter material surface, thereby remove MIB and Geosmin in tap water.
The invention has the advantages that:
(1) improved biological filter causes and smells material degradation capability is arranged the typical case, in the settling tank water outlet that research discovery MIB and Geosmin starting point concentration are 500ng/L, removal 67%(74% when EBCT is 20min) MIB(Geosmin), greatly reduced ozone and activated carbon dosage in subsequent technique, had advantages of that cost is low, effective.
(2) bacterial classification of screening is taken from the waterworks activated carbon filter, be easy to cultivate and to human body toxicological harmless effect, there are safe, economic characteristics, having very important significance aspect degraded removal blue-green algae and the microbial tap water MIB of unwrapping wire and Geosmin pollution problem.
The accompanying drawing explanation
Fig. 1 is the technical process that biological filter is inoculated.
Fig. 2 is that the technical process containing MIB and the water outlet of Geosmin settling tank is processed in biological filter.
Fig. 3 a and Fig. 3 b are the removal effect schematic diagram of single degradation bacteria to MIB and Geosmin.
Fig. 4 is the impact of the empty bed residence time on water outlet MIB and Geosmin concentration.
Fig. 5 is the impact of filter tank height on water outlet MIB and Geosmin concentration.
in figure:
1. reactor, 2. water tank, 3. blank assay post, 4. common sand filtration post, 5. improved sand filtration post, 6. blank assay post.
specific implementation method
below in conjunction with specific embodiment, technical scheme of the present invention is described further.
embodiment 1:
(1) preparation minimal medium: by the NH of 1.0g
4nO
3, the KH of 1.0g
2pO
4, the MgSO of 0.5g
47H
2the KCl of O and 0.2g is dissolved in the distilled water of 1000mL, regulate the pH value to 7.2-7.4, in temperature, be sterilizing 30min under 121 ° of C, with water system needle type filtration device, the MIB that is 20 μ g/L by the concentration of 1-100 μ L respectively under gnotobasis and Geosmin standardized solution add in aseptic 200mL minimal medium, obtain concentration and be respectively 100ng/L, 200ng/L, 500ng/L, 1 μ g/L, 2 μ g/L, 5 μ g/L and the MIB of 10 μ g/L and the minimal medium of Geosmin, standby;
(2) preparation LB solid medium: the NaCl of the yeast extract of the peptone of 10g, 5g and 10g is dissolved in the distilled water of 1000mL, the NaOH that is 1mol/L by concentration regulates pH value to 7.2, then adds the 15g agar powder, and heating is dissolved agar powder, then cool to room temperature, standby;
(3) preparation PBS damping fluid: by Na
2hPO
412H
2o is dissolved in preparation in distilled water and obtains the Na that concentration is 0.2 mol/L
2hPO
4, by NaH
2pO
42H
2o is dissolved in distilled water configuration, and to obtain concentration be 0.2 mol/L NaH
2pO
4; To prepare NaH
2pO
4, Na
2hPO
4after mixing according to the volume ratio of 19:81:200 with water, in temperature, be sterilizing 30min under 121 ° of C, the PBS damping fluid that to obtain concentration be 0.1mol/L, pH=7.4, be stored in dark cold place, standby;
(4) get the gac of biofilm development maturation in the 9th water factory's activated carbon filter of 5.0g Beijing, add 30mL 0.1mol/L PBS damping fluid concuss 10 min, be dissolved in the minimal medium of 200mL containing 100ng/L MIB and Geosmin and carry out selectivity cultivation enrichment with the absorption of the suction pipe after sterilizing 10mL bacterium liquid after standing for some time, be placed under constant-temperature shaking incubator (30 ° of C, 160rpm) condition and cultivate.
(5) after the substratum muddiness, according to 10% inoculum size, progressively be transferred in the minimal medium of 200ng/L, 500ng/L, 1 μ g/L, 2 μ g/L, 5 μ g/L and 10 μ g/L MIB and Geosmin respectively, the typical case is caused and smells the mass degradation bacterium and screened, extract the typical case and cause and smell the mass degradation bacterium.
(6) after each liquid culture stage end, with the aseptic technique method, draw the water sample that 1mL fully mixes, inject and fill the 9mL test tube of sterile saline, be mixed into the 1:10 diluent; The diluent 1mL that draws 1:10 annotates the people and fills in the test tube of 9mL sterile saline, is mixed into the 1:100 diluent.Be diluted to successively by the same method 1:10
3, 1:10
4, 1:10
5, 1:10
6, 1:10
7standby Deng diluent etc.So increase progressively dilution once, must change a 1mL sterilizing suction pipe.
(7) get different dilution water sample 0.2mL with the sterilizing suction pipe, inject respectively in the plate contain freezing solid substratum, with the glass scraper by bacteria suspension in the media surface coating evenly.Before use, the glass scraper need be through aseptically process, with after calcination sterilizing on spirit lamp immediately, place coolingly, be used alternatingly.Do a parallel inoculation during each check, separately with a plate, only pour into the LB solid medium as blank simultaneously.After bacterium liquid absorbs, the upset plate, make bottom surface upwards, is placed in ° C incubator of 30 ° of C ± 1 and cultivates 48h, with strain screening.
(8) adopt plate streak separation and purification degraded bacterial classification, obtaining micrococci, Flavobacterium, tyrothricin and pseudomonas is the MIB degradation bacteria, and golden Zymomonas mobilis, Sinorhizobium and Stenotrophomonas are the Geosmin degradation bacteria.
(10) sand grains after cleaning is inserted to Filter column.Adopt containing acquired typical case and cause and smell mass degradation bacterium inoculation filter bed, concrete grammar is as follows: will contain every kind of bacterium each 1 * 10
3the bacterium liquid of individual/mL be take 1mL/min(EBCT as 30min) flow through Filter column and circulate 4 days, make degradation bacteria can be attached to filter material surface; Adopt sterilized water to flow through the filter post in contrast with same flow velocity.
(11) inoculation liquid is emptying, Beijing of containing MIB and each 500ng/L of Geosmin the 9th each 25L of settling tank water outlet of water factory after 3 barrels of sterilizings is placed in respectively by reactor, with certain filtering velocity flow through blank, do not inoculate and inoculate sand filter, investigate the impact of EBCT.In Fig. 3, a is the removal effect of single culture to MIB in the settling tank water outlet, and b is the removal effect of single culture to Geosmin in the settling tank water outlet.In Fig. 3 a, 1 is blank, and 2 is the micrococci treatment effect, and 3 is the Flavobacterium treatment effect, and 4 is the tyrothricin treatment effect, and 5 is the pseudomonas treatment effect; In Fig. 3 b, 1 is blank, and 2 is golden Zymomonas mobilis treatment effect, and 3 is the Sinorhizobium treatment effect, and 4 is the Stenotrophomonas treatment effect; Fig. 3 shows that the bacterial classification that screening obtains all has removal effect preferably to MIB and Geosmin, and micrococci, Flavobacterium, tyrothricin and pseudomonas are respectively 98%, 96% to the clearance of MIB, 95% and 93%, and wherein the MIB of volatilization accounts for 25%; Golden Zymomonas mobilis, Sinorhizobium and Stenotrophomonas are respectively 85%, 82% and 83% to the clearance of Geosmin, and wherein the Geosmin of volatilization accounts for 35%.
In Fig. 3 b, in Fig. 4, being the removal effect of biological filter to MIB in the settling tank water outlet, is the removal effect of biological filter to Geosmin in the settling tank water outlet.In Fig. 4,1 ~ 3 represents respectively blank, common sand filtration post and the improved sand filtration post removal effect to MIB and Geosmin.Fig. 4 shows in the settling tank water outlet that MIB and Geosmin starting point concentration are 500ng/L, and filtrate height 550mm, when EBCT is 20min, 67%(74% can be removed in improved biological filter) MIB(Geosmin).
embodiment 2:
(1) screening obtains the bacterial classification of purifying
(2) by experiment in chamber experimental simulation water factory biological filter typical case in water is caused to the removal effect of smelling material and other index.Sand grains after cleaning is inserted to Filter column.Adopt containing acquired typical case and cause and smell mass degradation bacterium inoculation filter bed, concrete grammar is as follows: will contain every kind of bacterium each 1 * 10
3the bacterium liquid of individual/mL be take 1mL/min(EBCT as 20min) flow through Filter column and circulate 4 days, make degradation bacteria can be attached to filter material surface; Adopt sterilized water to flow through the filter post in contrast with same flow velocity.
(3) inoculation liquid is emptying, Beijing of containing MIB and each 500ng/L of Geosmin the 9th each 25L of settling tank water outlet of water factory after 3 barrels of sterilizings is placed in respectively by reactor, with certain filtering velocity flow through blank, do not inoculate and inoculate sand filter, investigate the impact of filtrate height.
In Fig. 5, being the removal effect of biological filter to MIB in the settling tank water outlet, is the removal effect of biological filter to Geosmin in the settling tank water outlet.In Fig. 5,1 and 2 represent respectively common sand filtration post and the improved sand filtration post removal effect to MIB and Geosmin.
Claims (1)
1. the method for MIB and Geosmin in water is removed in the inoculation biological filter
, it is characterized in that, specifically comprise the following steps:
step 1:
1) preparation minimal medium: by the NH of 0.8-1.2g
4nO
3, 0.8-1.2g KH
2pO
4, 0.4-0.6g MgSO
47H
2the KCl of O and 0.2g is dissolved in the distilled water of 1000mL, regulate the pH value to 7.2-7.4, in temperature, be sterilizing 30min under 121 ° of C, with water system needle type filtration device, the MIB that is 20 μ g/L by the concentration of 1-100 μ L under gnotobasis and Geosmin standardized solution add respectively in aseptic 200mL minimal medium, obtain concentration and be respectively 100ng/L, 200ng/L, 500ng/L, 1 μ g/L, 2 μ g/L, 5 μ g/L and the MIB of 10 μ g/L and the minimal medium of Geosmin, standby;
2) preparation LB solid medium: the NaCl of the yeast extract of the peptone of 5-10g, 3-5g and 8-10g is dissolved in the distilled water of 1000mL, the NaOH that is 1mol/L by concentration regulates the pH value to 7.2-7.4, then add the 10-15g agar powder, heating is dissolved agar powder, then cool to room temperature, standby;
3) preparation damping fluid: by Na
2hPO
412H
2o is dissolved in preparation in distilled water and obtains the Na that concentration is 0.2 mol/L
2hPO
4, by NaH
2pO
42H
2o is dissolved in distilled water configuration, and to obtain concentration be 0.2 mol/L NaH
2pO
4; To prepare NaH
2pO
4, Na
2hPO
4after mixing according to the volume ratio of 19:81:200 with water, in temperature, be sterilizing 30min under 121 ° of C, the PBS damping fluid that to obtain concentration be 0.1mol/L, pH=7.4, be stored in dark cold place, standby;
Step 2:
1) get the PBS damping fluid that the 30mL above-mentioned steps prepares, the gac that adds biofilm development maturation in a certain amount of activated carbon filter, concuss 10 min, standing for some time, sterilizing, obtain biomembranous PBS damping fluid, with suction pipe draw 10mL containing biomembranous PBS damping fluid, be dissolved in 200mL step 1 1) obtain carrying out selectivity in the minimal medium of MIB that concentration is 100ng/L and Geosmin and cultivate enrichment, being placed in constant-temperature shaking incubator is 30 ° of C in temperature, under the condition that rotating speed is 160rpm, cultivate, after the substratum muddiness, respectively according to 10% inoculum size progressively be transferred to step 1 1) 200ng/L that obtains, 500ng/L, 1 μ g/L, 2 μ g/L, 5 μ g/L, in the MIB of 10 μ g/L and the minimal medium of Geosmin, after each liquid culture stage finishes, draw with the aseptic technique method water sample that 1mL fully mixes, injection fills the 9mL test tube of sterile saline, be mixed into the 1:10 diluent, the diluent 1mL that draws 1:10 annotates the people and fills in the test tube of 9mL sterile saline, is mixed into the 1:100 diluent, is diluted to successively by the same method 1:10
3, 1:10
4, 1:10
5, 1:10
6, 1:10
7diluent, standby, get different dilution diluent 0.2mL with the sterilizing suction pipe, inject respectively in the plate contain freezing solid substratum, with the glass scraper by bacteria suspension in the media surface coating evenly, before use, the glass scraper need be through aseptically process, with after calcination sterilizing on spirit lamp immediately, place cooling, be used alternatingly, do a parallel inoculation during each check, separately with a plate, only pour into the LB solid medium as blank simultaneously, after bacterium liquid absorbs, the upset plate, make bottom surface upwards, be placed in ° C incubator of 30 ° of C ± 1 and cultivate 48h, adopt plate streak separation and purification degraded bacterial classification, obtaining micrococci, Flavobacterium, tyrothricin and pseudomonas is the MIB degradation bacteria, and golden Zymomonas mobilis, Sinorhizobium and Stenotrophomonas are the Geosmin degradation bacteria,
Step 3:
It is highly the Filter column of 100-550mm that sand grains after cleaning is inserted, and the concentration that will contain MIB degradation bacteria and Geosmin degradation bacteria is 1 * 10
3the bacterium liquid of individual/mL, at empty bed residence time 6-60min, flow through Filter column and circulate 4 days, makes MIB degradation bacteria and Geosmin degradation bacteria be attached to described filter material surface, thereby remove MIB and Geosmin in water.
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| CN112730650B (en) * | 2020-12-15 | 2022-08-09 | 湖北微谱技术有限公司 | Method for enriching ultra-trace organic matters in large-volume water |
| WO2023147659A1 (en) * | 2022-02-01 | 2023-08-10 | 683107 Alberta Ltd. | Compositions, systems, and methods for processing recirculating aquaculture water |
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