CN107828773A - A kind of preparation method of the domestication agent of the activated sludge of brine waste processing - Google Patents
A kind of preparation method of the domestication agent of the activated sludge of brine waste processing Download PDFInfo
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Abstract
The invention belongs to biochemical treatment of industrial wastewater field, and in particular to a kind of preparation method of the domestication agent of the activated sludge of brine waste processing.The present invention screens a kind of efficient Halophiles from butcher's meat, add the strain of thermophilic salt, improve microorganism group into, the activity of intensified response main body activated sludge, because of its special Mechanism of Salt-tolerant, suitable in hypersaline environment growth and degradation of organic substances, Halophiles both can directly absorb glycine betaine and tetrahydropyrimidine etc. from environment, can also voluntarily it be synthesized in the cell by absorbing its substrate, this mechanism can make microorganism resist the change of osmotic pressure in hypersaline environment, and then there is stronger adaptive faculty, improve salt resistance degree impact capacity.
Description
Technical field
The invention belongs to biochemical treatment of industrial wastewater field, and in particular to a kind of activated sludge of brine waste processing is tamed and dociled
Change method.
Background technology
High-salt wastewater refers to waste water of the inorganic salts mass fraction more than 1% in water, and its source includes：（1）Seawater utilization：Sea
Water processing, mariculture and waste water caused by seawater substitute discharge；（2）Industrial wastewater：Food, oil, chemical industry, printing and dyeing, papermaking,
The industries such as leather, pharmacy, fermentation produce high saliferous organic wastewater with high concentration extensively；（3）It is other：Ship ballast water, wastewater minimization
Caused brackish water（Percolate, reverse osmosis concentrated draining）, large-scale ships sanitary sewage.In coastal area, if can be by high salt
Organic matter and nutriment in waste water are reduced to as defined in discharge standard below limit value, and directly arranging sea will not make to ecological environment
Into big influence；In hinterland, it is necessary to while remove the inorganic salts in high-salt wastewater, organic matter and nutrients and could discharge,
Otherwise any biased processing or way to manage can all cause serious harm to ecological environment.Therefore, in coastal area, according to
Biochemical method and combinations thereof technique handles the pollutant thing in high-salt wastewater and nutrients to below limited value of discharge standard, can be straight
Run in and put, can largely reduce investment and the operating cost of high-salt wastewater processing；In hinterland, according to based on life
Change and combinations thereof technique and carry out desalination by after high-salt wastewater purified treatment, then using embrane method, evaporation or other techniques, reclaim
To inorganic salts slightly refine or do not do the refined Nacl that can be used as higher degree, thus can offset partial deionization investment and
Operating cost, and fundamentally avoid the possibility of the generation of solid waste, the transfer of pollutant and secondary pollution.
Activated sludge is one kind as accompanying by the microorganisms such as bacterium, fungi, protozoan, metazoa and algae and they
Organic substance, the general name of floc sludge that forms of inorganic substances, main application is the processing of sanitary sewage and industrial wastewater, tool
There is the floc sludge of bioactivity and certain degrading waste water ability.
Relative to physical chemistry method, biochemical treatment is normally more economical friendly, is sanitary sewage and process for treating industrial waste water
Primary selection；But traditional view and engineering practice are thought：High salt concentration has to the growth metabolism of common micro-organisms cell
There are suppression or toxic action, biochemical treatment efficiency and stability are all poor under hypersaline environment.Therefore, general biochemical method can not be direct
For high-salt wastewater processing, it is necessary to obtain salt-durable microbe or improve the salt resistance ability of activated sludge, high salt can be adapted to and given up
The needs of water biochemical treatment.At present, the main method of high-salt wastewater biochemical treatment includes：Salinity domestication, dilution is biochemical, selection is resistance to
The high biochemical process of salinity, the enhanced biological processing of inoculation Halophiles.
The general acclimation method of activated sludge is divided into according to strain is added（1）Add activated sludge, breeding to predetermined sedimentation
Than when, add actual waste water blast aeration, by being tamed step by step to full dose on a small quantity.（2）Specified strain is added, first puts small incubation into
After the breeding of case hyperalimentation fluid, then put into great Chi and breed the settling ratio for reaching required again.It is divided into according to culture domestication program：（1）
Synchronous culture domestication, i.e., with excrement water or sanitary sewage culture activated sludge while add industrial wastewater to be processed.（2）It is different
Step culture domestication, i.e., first use excrement water or sanitary sewage culture activated sludge, then the incremental mode of concentration gradient adds industry
Waste water, until activated sludge reaches required settling ratio.
Salinity domestication ordinary activated sludge can make it obtain certain salt resistance ability, but the salt tolerant energy obtained by domestication
Power is very limited, is usually no more than 3.5%, not more than 5%；Moreover, the original for the treatment effect that biochemical treatment system is played stably
Cause：Broken dependent on the Complex Ecological Systems of multiple-microorganism population composing and its interaction, salinity domestication in activated sludge process
The highly stable biological community structure that common non-salt-durable microbe is formed is broken, promotes activated sludge during salinity domestication
In a small number of weak halophilic microorganisms and salt-durable microbe obtain competitive advantage, but due to species very little, domestication the time it is often inadequate,
And be difficult to form stable species community structure, so as to cause, treatment effeciency is not high and effect is unstable.Tame the activity obtained
Sludge salt resistance ability is unstable, salt resistance degree impact capacity is poor；Tame time length, treatment effeciency declines under hypersaline environment.It is first dilute
The biochemical suppression and toxic action that high salt concentration can be avoided to microorganism is released again, but wastewater treatment investment and operating cost all increase
Add, be only applicable to the situation that water is small and there are other less salt wastewater sources to be diluted.
Therefore, for existing high-salt wastewater biochemical processing method the defects of and limitation, we are necessary to propose a kind of letter
The cultural method of single easy-operating activated sludge available for brine waste processing, it is social and relevant department to be adapted to this
Demand.
The content of the invention
The technical problems to be solved by the invention：It is low for the salt resistance ability of current common salinity acclimated activated sludge, resist
Salinity shock ability, the problem for the treatment of effeciency is low under hypersaline environment, there is provided a kind of activated sludge of brine waste processing
Domestication agent preparation method.
In order to solve the above technical problems, the present invention is using technical scheme as described below：
A kind of preparation method of the domestication agent of the activated sludge of brine waste processing, the preparation method comprise the following steps：
（1）Butcher's meat stripping and slicing is taken, mass fraction is added to be well mixed in 0.9% physiological saline, vibrates, stand, take supernatant,
Supernatant is diluted to 10 using sterilized water-5Dilution level, obtain bacterium solution, take dilution after Halophiles bacterium solution be seeded to Halophiles enrichment
Cultivated in fluid nutrient medium, repeat enrichment culture 2 times, obtain Halophiles enrichment culture liquid, the line of picking Halophiles enrichment culture liquid
It is seeded to Halophiles and isolates and purifies plating medium, cultivate, obtains Halophiles culture, the maximum bacterium colony in picking bacterium footpath, repeat to draw
Line culture 2 ~ 3 times, must purify Halophiles bacterium colony, and picking purifying Halophiles colony inoculation is trained into Halophiles enriched liquid culture medium
Support, Halophiles nutrient solution must be purified, centrifuge, take mycelium, dry, obtain Halophiles powder；
（2）Take sludge to add sterilized water, vibrate, stand, take supernatant, and supernatant is added in nitrifying bacteria enrichment culture medium and trained
Support, obtain enrichment culture bacterium solution, by physiological saline gradient dilution that enrichment culture bacterium solution mass fraction is 0.9% to 10-7Dilution
Level, obtains bacterium solution after dilution, and bacterium solution is coated on nitrifier isolation and purification culture base after taking dilution, cultivates, and it is dizzy that picking produces blueness
The colony inoculation of circle must purify nitrifier bacterium colony, picking purifying nitrification to nitrifier isolation and purification culture base line separation 2 ~ 3 times
Bacterium colony inoculation is into nitrifying bacteria enrichment culture medium, culture, must purify nitrification bacteria culture fluid, centrifugation, take mycelium, dry, obtain
Nitrifier powder；
（3）Peanut shell based biomass charcoal is dried, grinding is sieved after smashing to pieces, peanut shell base carbon powder is obtained, by peanut shell base powdered carbon
End is added in KOH solution, immersion, obtains peanut shell base charcoal soak, peanut shell base charcoal soak is placed in quartz boat, is put into baking
In case, design temperature is 600 ~ 800 DEG C, constant temperature activation, must activate peanut shell base charcoal, is with concentration by activation peanut shell base charcoal
0.1mol/L hydrochloric acid solution washing, obtains pickling peanut shell base charcoal, pickling peanut shell base charcoal is washed with deionized to pH to 7.0
± 0.2, dry, peanut shell base porous charcoal is made；
（4）By Halophiles powder, nitrifier powder, peanut shell base porous charcoal, trehalose, glycine betaine in mass ratio 3：5：15：5：2
It is well mixed, you can the domestication agent of the activated sludge as brine waste processing.
The step（1）Middle Halophiles extraction raw material butcher's meat is purchased from the market of farm produce；Halophiles isolates and purifies plating medium
Formula for according to the mass fraction, take 40 ~ 50 parts of NaCl, anhydrous 9 ~ 11 parts of MgSO4,2 ~ 3 parts of sodium citrate, 2 ~ 5 parts of KCl,
Anhydrous CaCl20.2 ~ 0.5 part, 10 ~ 15 parts of peptone, 2 ~ 5 parts of yeast extract, 20 ~ 25 parts of agar, 1000 parts of water, pH 7.5
± 0.2,121 DEG C of 15 min of sterilizing；Halophiles enriched liquid culture medium：Remove Halophiles and isolate and purify agar in plating medium
Composition, other components are constant.
The step（1）Middle butcher's meat specification is 0.5cm × 0.5cm block, and butcher's meat and mass fraction are 0.9% physiological saline
Mass ratio be 1：10；The volume ratio of supernatant and sterilized water is 1：9；Halophiles bacterium solution by volume 1 after dilution：8 are seeded to
In Halophiles enriched liquid culture medium, 30 ~ 35 DEG C, 180r/min cultivate 2 ~ 3 days；The condition of culture of Halophiles culture be 30 ~
35 DEG C are cultivated 2 ~ 3 days；The condition of culture for purifying Halophiles nutrient solution is cultivated 3 ~ 5 days for 30 ~ 35 DEG C.
The step（2）Middle denitrifying bacterium extraction raw material is derived from settled sludge in the high-salt wastewater near certain pickles factory；Nitre
Change bacterium enriched medium：As mass fraction, 12 ~ 15 parts of peptone, NaCl6 ~ 8 part, 1 ~ 2 part of KNO3, sodium succinate 2 ~ 3 are taken
Part, 1000 parts of water, pH values are 7.0 ± 0.5；Nitrifier isolation and purification culture base：As mass fraction, Na2HPO4 7 ~ 8 is taken
Part, KH2PO42 ~ 3 part, MgSO47H2O0.05 ~ 0.1 part, KNO31 ~ 2 part, 9 ~ 12 parts of sodium succinate, bromthymol blue 0.1 ~
0.2 part, 5 ~ 6 parts of absolute ethyl alcohol, 20 ~ 25 parts of agar, 1000 parts of water, pH value 7.0 ± 0.3.
The step（2）The mass ratio of middle sludge and sterilized water is 1：10；Supernatant by volume 1：9 add nitrifier richness
Collect in culture medium, 28 ~ 30 DEG C of 18 ~ 24h of culture；Purifying nitrification bacteria culture fluid condition of culture is 30 ~ 35 DEG C and cultivated 2 ~ 3 days.
The step（3）Middle peanut hull biologic matter charcoal is bought in Henan Sanli Ltd.；Peanut shell base carbon powder and KOH solution
Mass ratio be 2：1, soak 24 ~ 36h.
The step（4）Middle Halophiles powder, nitrifier powder, peanut shell base porous charcoal, trehalose, the quality of glycine betaine
Than for 3：5：15：5：2.
Compared with other method, advantageous effects are the present invention：
（1）The present invention screens a kind of efficient Halophiles from butcher's meat, adds the strain of thermophilic salt, improves microorganism group into enhancing is anti-
The activity of main body activated sludge is answered, because of its special Mechanism of Salt-tolerant, suitable in hypersaline environment growth and degradation of organic substances, Halophiles
Both glycine betaine and tetrahydropyrimidine etc. can be directly absorbed from environment, can also voluntarily be closed in the cell by absorbing its substrate
Into this mechanism can make microorganism resist the change of osmotic pressure in hypersaline environment, and then have stronger adaptive faculty, improve anti-
Salinity shock ability；
（2）High-salt wastewater processing in, salinity height cause the dehydrogenase activity of denitrogenation strain reduces, cell plasmolysis, grow
Metabolism is suppressed, and this make it that not only nitric efficiency reduces conventional biological treatment, and organic matter can not be degraded very well,
Treatment effect can not meet the requirement of wastewater discharge standard, and the present invention screens efficient nitrification from high-salt wastewater in settled sludge
Bacterium, and this special efficient bacterium is invested in the activated sludge of high slat-containing wastewater, by acclimated activated sludge, can reduce
Ammonia-nitrogen content in high-salt wastewater, improve the nitric efficiency for high-salt wastewater；
（3）The present invention makes porous carbon material as thalline appendix matrix from peanut hull biologic matter charcoal, in porous carbon structure
Macropore can regard ion buffer so that diffusion path of the salt ion in duct greatly shortens, so as to be advantageous to salt ion
Rapid osmotic makes appendix act on high-salt wastewater in thalline therein, meso-hole structure causes porous to the internal gutter of porous charcoal
Carbon Materials have higher specific surface area, at the same time reduce internal resistance consumption when salt ion flows in the solution；
（4）Present invention addition trehalose and glycine betaine are highly water soluble the characteristics of these small-molecule substances as external source protective agent
Property, without electrostatic charge, high concentration accumulation in the cell does not interfere with the normal physiological work(of large biological molecule such as protein and nucleic acid
Can, but can be with the osmotic pressure outside statocyte, when microbial inoculum processing high-salt wastewater is impacted by salinity, Microbial Communities in Activated Sludge
Cell can quickly absorb the glycine betaine of addition and balance extraneous osmotic pressure, so as to shorten adaptation of the activated sludge to hypersaline environment
Phase, mitigate murder by poisoning of the salinity to activated sludge.
Brief description of the drawings
Fig. 1 is activated sludge salt tolerant implementations table.
Embodiment
Peanut hull biologic matter charcoal：Buy in Henan Sanli Ltd..
Halophiles extracts raw material：Butcher's meat, it is purchased from the market of farm produce.
Denitrifying bacterium extracts raw material：It is derived from settled sludge in the high-salt wastewater near certain pickles factory.
Halophiles isolates and purifies plating medium：According to the mass fraction, 40 ~ 50 parts of NaCl, anhydrous MgSO are taken4 9 ~ 11 parts,
2 ~ 3 parts of sodium citrate, 2 ~ 5 parts of KCl, anhydrous CaCl20.2 ~ 0.5 part, 10 ~ 15 parts of peptone, 2 ~ 5 parts of yeast extract, fine jade
20 ~ 25 parts of fat, 1000 parts of water, 7.5 ± 0.2,121 DEG C of 15 min of sterilizing of pH.
Halophiles enriched liquid culture medium：Remove Halophiles and isolate and purify agar composition in plating medium, other components
It is constant.
Nitrifying bacteria enrichment culture medium：As mass fraction, 12 ~ 15 parts of peptone, NaCl6 ~ 8 part, KNO are taken31 ~ 2 part, fourth
2 ~ 3 parts of diacid sodium, 1000 parts of water, pH values are 7.0 ± 0.5.
Nitrifier isolation and purification culture base：As mass fraction, Na is taken2HPO47 ~ 8 parts, KH2PO42 ~ 3 parts, MgSO4·
7H2O0.05 ~ 0.1 part, KNO31 ~ 2 part, 9 ~ 12 parts of sodium succinate, 0.1 ~ 0.2 part of bromthymol blue, 5 ~ 6 parts of absolute ethyl alcohol, agar
20 ~ 25 parts, 1000 parts of water, pH value 7.0 ± 0.3.
A kind of acclimation method of the activated sludge of brine waste processing, comprises the following steps：
（5）Take butcher's meat be cut into 0.5cm × 0.5cm bulk, in mass ratio 1：10 are added to mass fraction as in 0.9% physiological saline
It is well mixed, about 20min is vibrated, disperses cell, stands 5 ~ 10min, takes supernatant by volume 1：9 mix with sterilized water
It is even, and it is diluted to 10-5Dilution level, obtain bacterium solution, take Halophiles bacterium solution by volume 1 after dilution：8 are seeded to Halophiles pregnant solution
In body culture medium, 30 ~ 35 DEG C, 180r/min cultivate 2 ~ 3 days, repeat enrichment culture 2 times, obtain Halophiles enrichment culture liquid, picking
Halophiles enrichment culture liquid streak inoculation to Halophiles isolates and purifies plating medium, and 30 ~ 35 DEG C are cultivated 2 ~ 3 days, obtain Halophiles
Culture, the maximum bacterium colony in picking bacterium footpath, line culture 2 ~ 3 times is repeated, Halophiles bacterium colony, picking purifying Halophiles bacterium must be purified
Fall to be seeded in Halophiles enriched liquid culture medium, 30 ~ 35 DEG C are cultivated 3 ~ 5 days, must purify Halophiles nutrient solution, are centrifuged, are taken bacterium
Filament, dry, obtain Halophiles powder；
（6）Take sludge in mass ratio 1：10 add sterilized waters, vibrate 20min, stand, take supernatant, and by volume 1：9 to supernatant
Nitrifying bacteria enrichment culture medium is added in liquid, 28 ~ 30 DEG C of 18 ~ 24h of culture, enrichment culture bacterium solution is obtained, by enrichment culture bacterium solution matter
Physiological saline gradient dilution that fraction is 0.9% is measured to 10-7Dilution level, obtain bacterium solution after dilution, take dilution after bacterium solution be coated on
Nitrifier isolation and purification culture base, 28 ~ 33 DEG C are cultivated 2 ~ 3 days, and colony inoculation to the nitrifier that picking produces blue halos separates
Pure medium line separation 2 ~ 3 times, must purify nitrifier bacterium colony, and picking purifies nitrifier colony inoculation to nitrifying bacteria enrichment and trained
Support in base, 30 ~ 35 DEG C are cultivated 2 ~ 3 days, must be purified nitrification bacteria culture fluid, centrifugation, be taken mycelium, dry, obtain nitrifier powder；
（7）Peanut shell based biomass charcoal is dried at 100 ~ 105 DEG C, grinding crosses 200 mesh sieves after smashing to pieces, obtains peanut shell base powdered carbon
End, by peanut shell base carbon powder in mass ratio 2：1 adds in KOH solution, soaks 24 ~ 36h, obtains peanut shell base charcoal soak, will
Peanut shell base charcoal soak is placed in quartz boat, oven temperature is risen into 600 ~ 800 DEG C with 10 DEG C/min speed, constant temperature activation
30 ~ 120min, peanut shell base charcoal must be activated, the hydrochloric acid solution that peanut shell base charcoal is 0.1mol/L with concentration will be activated and washed, obtained
Pickling peanut shell base charcoal, pickling peanut shell base charcoal is washed with deionized to pH to 7.0 ± 0.2, dries, peanut shell Quito is made
Hole charcoal；
（8）By Halophiles powder, nitrifier powder, peanut shell base porous charcoal, trehalose, glycine betaine in mass ratio 3：5：15：5：2
It is well mixed, you can the domestication agent of the activated sludge as brine waste processing.
Embodiment 1
Peanut hull biologic matter charcoal：Buy in Henan Sanli Ltd..
Halophiles extracts raw material：Butcher's meat, it is purchased from the market of farm produce.
Denitrifying bacterium extracts raw material：It is derived from settled sludge in the high-salt wastewater near certain pickles factory.
Halophiles isolates and purifies plating medium：According to the mass fraction, 40 parts of NaCl, anhydrous MgSO are taken49 parts, lemon
Sour 2 parts of sodium, 2 parts of KCl, anhydrous CaCl20.2 part, 10 parts of peptone, 2 parts of yeast extract, 20 parts of agar, 1000 parts of water, pH
7.5 ± 0.2,121 DEG C of 15 min of sterilizing.
Halophiles enriched liquid culture medium：Remove Halophiles and isolate and purify agar composition in plating medium, other components
It is constant.
Nitrifying bacteria enrichment culture medium：As mass fraction, 12 parts of peptone, NaCl6 parts, KNO are taken31 part, sodium succinate 2
Part, 1000 parts of water, pH values are 7.0 ± 0.5.
Nitrifier isolation and purification culture base：As mass fraction, Na is taken2HPO47 parts, KH2PO42 parts, MgSO4·
7H2O0.05 parts, KNO31 part, 9 parts of sodium succinate, 0.1 part of bromthymol blue, 5 parts of absolute ethyl alcohol, 20 parts of agar, 1000 parts of water,
PH value 7.0 ± 0.3.
A kind of acclimation method of the activated sludge of brine waste processing, comprises the following steps：
（9）Take butcher's meat be cut into 0.5cm × 0.5cm bulk, in mass ratio 1：10 are added to mass fraction as in 0.9% physiological saline
It is well mixed, about 20min is vibrated, disperses cell, stands 5min, takes supernatant by volume 1：9 are well mixed with sterilized water,
And it is diluted to 10-5Dilution level, obtain bacterium solution, take Halophiles bacterium solution by volume 1 after dilution：8 are seeded to Halophiles enriched liquid
In culture medium, 30 DEG C, 180r/min cultivate 2 days, repeat enrichment culture 2 times, obtain Halophiles enrichment culture liquid, picking Halophiles is rich
Collection nutrient solution streak inoculation to Halophiles isolates and purifies plating medium, and 30 DEG C are cultivated 2 days, obtain Halophiles culture, picking bacterium
The maximum bacterium colony in footpath, line culture 2 times is repeated, Halophiles bacterium colony must be purified, picking purifies Halophiles colony inoculation to Halophiles
In enriched liquid culture medium, 30 DEG C are cultivated 3 days, must purify Halophiles nutrient solution, are centrifuged, are taken mycelium, are dried, are obtained Halophiles powder
End；
（10）Take sludge in mass ratio 1：10 add sterilized waters, vibrate 20min, stand, take supernatant, and by volume 1：9 is upward
Nitrifying bacteria enrichment culture medium is added in clear liquid, 28 DEG C of culture 18h, obtains enrichment culture bacterium solution, by enrichment culture bacterium solution quality point
The physiological saline gradient dilution for being 0.9% is counted to 10-7Dilution level, obtain bacterium solution after dilution, take dilution after bacterium solution be coated on nitrification
Bacterium isolation and purification culture base, 28 DEG C are cultivated 2 days, and picking produces the colony inoculation of blue halos to nitrifier isolation and purification culture base
Line separation 2 times, nitrifier bacterium colony must be purified, picking purifies nitrifier colony inoculation into nitrifying bacteria enrichment culture medium, 30 DEG C
Culture 2 days, nitrification bacteria culture fluid must be purified, centrifugation, takes mycelium, dries, obtains nitrifier powder；
（11）Peanut shell based biomass charcoal to be dried at 100 DEG C, grinding crosses 200 mesh sieves after smashing to pieces, obtains peanut shell base carbon powder,
By peanut shell base carbon powder in mass ratio 2：1 adds in KOH solution, soaks 24h, peanut shell base charcoal soak is obtained, by peanut shell
Base charcoal soak is placed in quartz boat, and oven temperature is risen into 600 DEG C with 10 DEG C/min speed, constant temperature activation 30min, obtains work
Change peanut shell base charcoal, the hydrochloric acid solution that peanut shell base charcoal is 0.1mol/L with concentration will be activated and washed, obtain pickling peanut shell base charcoal,
Pickling peanut shell base charcoal is washed with deionized to pH to 7.0 ± 0.2, dries, peanut shell base porous charcoal is made；
By Halophiles powder, nitrifier powder, peanut shell base porous charcoal, trehalose, glycine betaine in mass ratio 3：5：15：5：2 is mixed
Close uniform, you can the domestication agent of the activated sludge as brine waste processing.
Embodiment 2
Peanut hull biologic matter charcoal：Buy in Henan Sanli Ltd..
Halophiles extracts raw material：Butcher's meat, it is purchased from the market of farm produce.
Denitrifying bacterium extracts raw material：It is derived from settled sludge in the high-salt wastewater near certain pickles factory.
Halophiles isolates and purifies plating medium：According to the mass fraction, 45 parts of NaCl, anhydrous MgSO are taken4 10 parts, lemon
Sour 2 parts of sodium, 3 parts of KCl, anhydrous CaCl20.3 part, 13 parts of peptone, 3 parts of yeast extract, 23 parts of agar, 1000 parts of water, pH
7.5 ± 0.2,121 DEG C of 15 min of sterilizing.
Halophiles enriched liquid culture medium：Remove Halophiles and isolate and purify agar composition in plating medium, other components
It is constant.
Nitrifying bacteria enrichment culture medium：As mass fraction, 13 parts of peptone, NaCl7 parts, KNO are taken31 part, sodium succinate 2
Part, 1000 parts of water, pH values are 7.0 ± 0.5.
Nitrifier isolation and purification culture base：As mass fraction, Na is taken2HPO47 parts, KH2PO42 parts, MgSO4·
7H2O0.07 parts, KNO31 part, 10 parts of sodium succinate, 0.1 part of bromthymol blue, 5 parts of absolute ethyl alcohol, 22 parts of agar, 1000 parts of water,
PH value 7.0 ± 0.3.
A kind of acclimation method of the activated sludge of brine waste processing, comprises the following steps：
（12）Take butcher's meat be cut into 0.5cm × 0.5cm bulk, in mass ratio 1：10 are added to mass fraction as in 0.9% physiological saline
It is well mixed, about 20min is vibrated, disperses cell, stands 8min, takes supernatant by volume 1：9 are well mixed with sterilized water,
And it is diluted to 10-5Dilution level, obtain bacterium solution, take Halophiles bacterium solution by volume 1 after dilution：8 are seeded to Halophiles enriched liquid
In culture medium, 33 DEG C, 180r/min cultivate 2 days, repeat enrichment culture 2 times, obtain Halophiles enrichment culture liquid, picking Halophiles is rich
Collection nutrient solution streak inoculation to Halophiles isolates and purifies plating medium, and 33 DEG C are cultivated 2 days, obtain Halophiles culture, picking bacterium
The maximum bacterium colony in footpath, line culture 2 times is repeated, Halophiles bacterium colony must be purified, picking purifies Halophiles colony inoculation to Halophiles
In enriched liquid culture medium, 33 DEG C are cultivated 4 days, must purify Halophiles nutrient solution, are centrifuged, are taken mycelium, are dried, are obtained Halophiles powder
End；
（13）Take sludge in mass ratio 1：10 add sterilized waters, vibrate 20min, stand, take supernatant, and by volume 1：9 is upward
Nitrifying bacteria enrichment culture medium is added in clear liquid, 29 DEG C of culture 20h, obtains enrichment culture bacterium solution, by enrichment culture bacterium solution quality point
The physiological saline gradient dilution for being 0.9% is counted to 10-7Dilution level, obtain bacterium solution after dilution, take dilution after bacterium solution be coated on nitrification
Bacterium isolation and purification culture base, 30 DEG C are cultivated 2 days, and picking produces the colony inoculation of blue halos to nitrifier isolation and purification culture base
Line separation 2 times, nitrifier bacterium colony must be purified, picking purifies nitrifier colony inoculation into nitrifying bacteria enrichment culture medium, 33 DEG C
Culture 2 days, nitrification bacteria culture fluid must be purified, centrifugation, takes mycelium, dries, obtains nitrifier powder；
（14）Peanut shell based biomass charcoal to be dried at 101 DEG C, grinding crosses 200 mesh sieves after smashing to pieces, obtains peanut shell base carbon powder,
By peanut shell base carbon powder in mass ratio 2：1 adds in KOH solution, soaks 30h, peanut shell base charcoal soak is obtained, by peanut shell
Base charcoal soak is placed in quartz boat, and oven temperature is risen into 700 DEG C with 10 DEG C/min speed, constant temperature activation 70min, obtains work
Change peanut shell base charcoal, the hydrochloric acid solution that peanut shell base charcoal is 0.1mol/L with concentration will be activated and washed, obtain pickling peanut shell base charcoal,
Pickling peanut shell base charcoal is washed with deionized to pH to 7.0 ± 0.2, dries, peanut shell base porous charcoal is made；
By Halophiles powder, nitrifier powder, peanut shell base porous charcoal, trehalose, glycine betaine in mass ratio 3：5：15：5：2 is mixed
Close uniform, you can the domestication agent of the activated sludge as brine waste processing.
Embodiment 3
Peanut hull biologic matter charcoal：Buy in Henan Sanli Ltd..
Halophiles extracts raw material：Butcher's meat, it is purchased from the market of farm produce.
Denitrifying bacterium extracts raw material：It is derived from settled sludge in the high-salt wastewater near certain pickles factory.
Halophiles isolates and purifies plating medium：According to the mass fraction, 50 parts of NaCl, anhydrous MgSO are taken411 parts, lemon
Sour 3 parts of sodium, KCl5 parts, anhydrous CaCl20.5 part, 15 parts of peptone, 5 parts of yeast extract, 25 parts of agar, 1000 parts of water, pH
7.5 ± 0.2,121 DEG C of 15 min of sterilizing.
Halophiles enriched liquid culture medium：Remove Halophiles and isolate and purify agar composition in plating medium, other components
It is constant.
Nitrifying bacteria enrichment culture medium：As mass fraction, 15 parts of peptone, 8 parts of NaCl, KNO are taken32 parts, sodium succinate
3 parts, 1000 parts of water, pH values are 7.0 ± 0.5.
Nitrifier isolation and purification culture base：As mass fraction, Na is taken2HPO48 parts, KH2PO43 parts, MgSO4·
7H2O0.1 parts, KNO32 parts, 12 parts of sodium succinate, 0.2 part of bromthymol blue, 6 parts of absolute ethyl alcohol, 25 parts of agar, 1000 parts of water,
PH value 7.0 ± 0.3.
A kind of acclimation method of the activated sludge of brine waste processing, comprises the following steps：
（15）Take butcher's meat be cut into 0.5cm × 0.5cm bulk, in mass ratio 1：10 are added to mass fraction as in 0.9% physiological saline
It is well mixed, about 20min is vibrated, disperses cell, stands 10min, takes supernatant by volume 1：9 mix with sterilized water
It is even, and it is diluted to 10-5Dilution level, obtain bacterium solution, take Halophiles bacterium solution by volume 1 after dilution：8 are seeded to Halophiles pregnant solution
In body culture medium, 35 DEG C, 180r/min cultivate 3 days, repeat enrichment culture 2 times, obtain Halophiles enrichment culture liquid, picking Halophiles
Enrichment culture liquid streak inoculation to Halophiles isolates and purifies plating medium, and 35 DEG C are cultivated 3 days, obtain Halophiles culture, picking
The maximum bacterium colony in bacterium footpath, line culture 3 times is repeated, Halophiles bacterium colony must be purified, picking purifies Halophiles colony inoculation to thermophilic salt
In bacterium enriched liquid culture medium, 35 DEG C are cultivated 5 days, must purify Halophiles nutrient solution, are centrifuged, are taken mycelium, are dried, are obtained Halophiles
Powder；
（16）Take sludge in mass ratio 1：10 add sterilized waters, vibrate 20min, stand, take supernatant, and by volume 1：9 is upward
Nitrifying bacteria enrichment culture medium is added in clear liquid, 30 DEG C of culture 24h, obtains enrichment culture bacterium solution, by enrichment culture bacterium solution quality point
The physiological saline gradient dilution for being 0.9% is counted to 10-7Dilution level, obtain bacterium solution after dilution, take dilution after bacterium solution be coated on nitrification
Bacterium isolation and purification culture base, 33 DEG C are cultivated 3 days, and picking produces the colony inoculation of blue halos to nitrifier isolation and purification culture base
Line separation 3 times, nitrifier bacterium colony must be purified, picking purifies nitrifier colony inoculation into nitrifying bacteria enrichment culture medium, 35 DEG C
Culture 3 days, nitrification bacteria culture fluid must be purified, centrifugation, takes mycelium, dries, obtains nitrifier powder；
（17）Peanut shell based biomass charcoal to be dried at 105 DEG C, grinding crosses 200 mesh sieves after smashing to pieces, obtains peanut shell base carbon powder,
By peanut shell base carbon powder in mass ratio 2：1 adds in KOH solution, soaks 36h, peanut shell base charcoal soak is obtained, by peanut shell
Base charcoal soak is placed in quartz boat, and oven temperature is risen into 800 DEG C with 10 DEG C/min speed, constant temperature activation 120min, obtained
Peanut shell base charcoal is activated, the hydrochloric acid solution that peanut shell base charcoal is 0.1mol/L with concentration will be activated and washed, obtain pickling peanut shell base
Charcoal, pickling peanut shell base charcoal is washed with deionized to pH to 7.0 ± 0.2, dries, peanut shell base porous charcoal is made；
By Halophiles powder, nitrifier powder, peanut shell base porous charcoal, trehalose, glycine betaine in mass ratio 3：5：15：5：2 is mixed
Close uniform, you can the domestication agent of the activated sludge as brine waste processing.
Comparative example：The activated sludge of Shanghai Bioisystech Co., Ltd production.
Method：The activated sludge prepared by the embodiment and comparative example of equivalent is taken, the brine waste of quality such as places it in
In, the salinity of waste water is improved successively, detects the activity of sludge, and the salt tolerance of sludge is characterized with this.Examined according to GB13456-92
Survey the COD clearances of activated sludge.
The specific detection case of activated sludge such as table 1
Table 1
| Detection project | Embodiment 1 | Embodiment 2 | Embodiment 3 | Comparative example |
| COD clearances（%） | 70.66 | 71.21 | 73.98 | 56.12 |
The specific detection case of activated sludge salt tolerance is shown in Figure of description Fig. 1.
From the foregoing, it will be observed that the activated sludge salt resistant character prepared by the present invention is good, COD clearances are high, are a kind of safety and height
The activated sludge of effect.
Claims (7)
1. a kind of preparation method of the domestication agent of the activated sludge of brine waste processing, it is characterised in that the preparation method includes
Following steps：
Butcher's meat stripping and slicing is taken, mass fraction is added to be well mixed in 0.9% physiological saline, vibrates, stand, take supernatant, use
Supernatant is diluted to 10 by sterilized water-5Dilution level, obtain bacterium solution, take dilution after Halophiles bacterium solution be seeded to Halophiles enriched liquid
Cultivated in culture medium, repeat enrichment culture 2 times, obtain Halophiles enrichment culture liquid, picking Halophiles enrichment culture liquid streak inoculation
Plating medium is isolated and purified to Halophiles, is cultivated, Halophiles culture is obtained, the maximum bacterium colony in picking bacterium footpath, repeats line training
To support 2 ~ 3 times, Halophiles bacterium colony must be purified, picking purifying Halophiles colony inoculation is cultivated into Halophiles enriched liquid culture medium,
Halophiles nutrient solution must be purified, centrifuges, takes mycelium, dries, obtains Halophiles powder；
Take sludge to add sterilized water, vibrate, stand, take supernatant, and supernatant is added in nitrifying bacteria enrichment culture medium and cultivated,
Enrichment culture bacterium solution, by physiological saline gradient dilution that enrichment culture bacterium solution mass fraction is 0.9% to 10-7Dilution level,
Bacterium solution after dilution, bacterium solution is coated on nitrifier isolation and purification culture base after taking dilution, cultivates, and picking produces blue halos
Colony inoculation must purify nitrifier bacterium colony, picking purifying nitrifier bacterium to nitrifier isolation and purification culture base line separation 2 ~ 3 times
Fall to be seeded in nitrifying bacteria enrichment culture medium, cultivate, nitrification bacteria culture fluid must be purified, centrifugation, take mycelium, dry, must nitrify
Bacterium powder end；
Peanut shell based biomass charcoal is dried, grinding is sieved after smashing to pieces, obtains peanut shell base carbon powder, peanut shell base carbon powder is added
Enter in KOH solution, soak, obtain peanut shell base charcoal soak, peanut shell base charcoal soak is placed in quartz boat, is put into baking oven
In, design temperature is 600 ~ 800 DEG C, constant temperature activation, must activate peanut shell base charcoal, is with concentration by activation peanut shell base charcoal
0.1mol/L hydrochloric acid solution washing, obtains pickling peanut shell base charcoal, pickling peanut shell base charcoal is washed with deionized to pH to 7.0
± 0.2, dry, peanut shell base porous charcoal is made；
By Halophiles powder, nitrifier powder, peanut shell base porous charcoal, trehalose, glycine betaine in mass ratio 3：5：15：5：2 is mixed
Close uniform, you can the domestication agent of the activated sludge as brine waste processing.
2. the preparation method of the domestication agent of the activated sludge of brine waste processing according to claim 1, it is characterised in that
The step（1）Middle Halophiles extraction raw material butcher's meat is purchased from the market of farm produce；The formula that Halophiles isolates and purifies plating medium is
According to the mass fraction, 40 ~ 50 parts of NaCl, anhydrous MgSO are taken49 ~ 11 parts, 2 ~ 3 parts of sodium citrate, 2 ~ 5 parts of KCl, anhydrous CaCl2
0.2 ~ 0.5 part, 10 ~ 15 parts of peptone, 2 ~ 5 parts of yeast extract, 20 ~ 25 parts of agar, 1000 parts of water, pH 7.5 ± 0.2,121
DEG C sterilizing 15 min；Halophiles enriched liquid culture medium：Remove Halophiles and isolate and purify agar composition in plating medium, other
Component is constant.
3. the preparation method of the domestication agent of the activated sludge of brine waste processing according to claim 1, it is characterised in that
The step（1）Middle butcher's meat specification is 0.5cm × 0.5cm block, butcher's meat and the mass ratio that mass fraction is 0.9% physiological saline
For 1：10；The volume ratio of supernatant and sterilized water is 1：9；Halophiles bacterium solution by volume 1 after dilution：8 are seeded to Halophiles richness
Collect fluid nutrient medium in, 30 ~ 35 DEG C, 180r/min cultivate 2 ~ 3 days；The condition of culture of Halophiles culture is 30 ~ 35 DEG C of cultures 2
~ 3 days；The condition of culture for purifying Halophiles nutrient solution is cultivated 3 ~ 5 days for 30 ~ 35 DEG C.
4. the preparation method of the domestication agent of the activated sludge of brine waste processing according to claim 1, it is characterised in that
The step（2）Middle denitrifying bacterium extraction raw material is derived from settled sludge in the high-salt wastewater near certain pickles factory；Nitrifying bacteria enrichment
Culture medium：As mass fraction, 12 ~ 15 parts of peptone, NaCl6 ~ 8 part, KNO are taken31 ~ 2 part, 2 ~ 3 parts of sodium succinate, water 1000
Part, pH values are 7.0 ± 0.5；Nitrifier isolation and purification culture base：As mass fraction, Na is taken2HPO47 ~ 8 parts, KH2PO42~3
Part, MgSO4·7H2O0.05 ~ 0.1 part, KNO31 ~ 2 part, 9 ~ 12 parts of sodium succinate, 0.1 ~ 0.2 part of bromthymol blue, absolute ethyl alcohol
5 ~ 6 parts, 20 ~ 25 parts of agar, 1000 parts of water, pH value 7.0 ± 0.3.
5. the preparation method of the domestication agent of the activated sludge of brine waste processing according to claim 1, it is characterised in that
The step（2）The mass ratio of middle sludge and sterilized water is 1：10；Supernatant by volume 1：9 add nitrifying bacteria enrichment culture medium
In, 28 ~ 30 DEG C of 18 ~ 24h of culture；Purifying nitrification bacteria culture fluid condition of culture is 30 ~ 35 DEG C and cultivated 2 ~ 3 days.
6. the preparation method of the domestication agent of the activated sludge of brine waste processing according to claim 1, it is characterised in that
The step（3）Middle peanut hull biologic matter charcoal is bought in Henan Sanli Ltd.；The mass ratio of peanut shell base carbon powder and KOH solution
For 2：1, soak 24 ~ 36h.
7. the preparation method of the domestication agent of the activated sludge of brine waste processing according to claim 1, it is characterised in that
The step（4）Middle Halophiles powder, nitrifier powder, peanut shell base porous charcoal, trehalose, the mass ratio of glycine betaine are 3：5：
15：5：2.
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| CN110386728A (en) * | 2019-07-30 | 2019-10-29 | 宜兴国际环保城科技发展有限公司 | A kind of tubular type free-radical oxidation handles the integral process of the high COD industrial wastewater of high salinity |
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