CN105062945A - Strain domesticating kit for degrading COD in coal-to-oil wastewater and domesticating method - Google Patents
Strain domesticating kit for degrading COD in coal-to-oil wastewater and domesticating method Download PDFInfo
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Abstract
The invention discloses a strain domesticating kit for degrading COD in coal-to-oil wastewater and a domesticating method. The kit comprises an enriching tube, a domestication prefabrication tube A and a domestication prefabrication tube B. The domesticating method comprises the steps of anoxic domestication and aerobic domestication. The kit is formed on the basis that a great number of coal-to-oil wastewater samples are collected in a laboratory within the whole nation, the components of the wastewater are analyzed and a domestication culture medium capable of simulating true wastewater is prepared through experiments. The domestication culture medium has relatively high adaptability; after strains domesticated by using the kit are added into various types of coal-to-oil wastewater, the COD degradation capability is improved, and the wastewater treatment starting time is greatly shortened; the kit can be used for simulating anoxic and aerobic biological processes and can be applied to product optimization of a coal-to-oil wastewater treatment agent and rapid domestication of sludge before the operation of a wastewater treatment plant.
Description
Technical field:
The invention belongs to a kind of strain domestication test kit and acclimation method, particularly one is for coal liquifaction waste water COD tamed strain test kit and the acclimation method of degrading.
Background technology:
Coal liquifaction take coal as raw material, produced a technology of oil product and petrochemicals, comprise DCL/Direct coal liquefaction and ICL for Indirect Coal Liquefaction two kinds of technological lines by chiral process.China's coal reserves account for 36% of world's total storage, account for more than 70% of China's energy total amount, and therefore in China, coal liquifaction is the leading of fuel chemical industry.
The feature of the oily(waste)water produced in coal liquifaction production process is: the toxic substance concentration such as polycyclic aromatic hydrocarbons and ramification of benzene, phenol, sulphur are high, and biodegradability is poor, is the more unmanageable waste water of one.In comprehensive wastewater, COD is unusual at about 1000-5000mg/L according to sewage treatment process.The harm of oily(waste)water is mainly manifested in: the film floating that oily substance is formed is on the water surface, oxygen in air just can not be dissolved in the water, dissolved oxygen in water will reduce, to cause in water body planktonic organism etc. dead because of anoxic, the photosynthesis of waterplant is also restricted, thus the self-purification of water body will be affected, water quality also can foul, the utility value of water resources also can be destroyed, and in oil, the existence of some lower boiling arene compounds can cause the generation of human cancer, thus affect HUMAN HEALTH.
The dominating process route of current domestic process coal liquifaction waste water is acted on " pre-treatment+biochemical treatment+advanced treatment " substantially, and wherein biochemical treatment is the important stage of sewage treatment process.By visiting domestic multiple coal liquifaction enterprise, at present for pretreated coal liquifaction waste water, generally adopt anoxic, aerobe method process (A/O technique) on the spot.Microorganism is the core of biological wastewater treatment process, governs the effect of sewage disposal.Sewage Plant initial start stage or operation be subject to impact need again to resume operation time, the domestic method generally adopted is in sewage treatment facility, add mud and nutritive substance, or add existing special microorganism microbial inoculum on market, reach the object that sewage treatment facility is normally run.For the process of coal liquifaction waste water, the effect adding mud and microbial inoculum is subject to the restriction of following several respects problem: one, contain the multiple material to the toxic effect of microorganism in coal liquifaction waste water, be added to the mud in sewage or the part microorganism in bacterial classification easily suppress by the toxicant in waste water, for head it off often needs to tame for a long time before adding mud or bacterial classification; Two, have the hardly degraded organic substances such as a large amount of oils in coal liquifaction waste water, these materials are difficult to utilized by microorganism and degrade, and address this problem and usually need the domestication of the appropriate design of technique and mud or microbial inoculum jointly to solve; Three, compared with common sewage, the start time of coal liquifaction waste water disposal facility is longer, needs the domestication of longer time.
Summary of the invention:
The object of the invention is to overcome the above problems, provide a kind of strain domestication test kit and acclimation method, the bacterial classification after the domestication of this test kit is rendered in coal liquifaction waste water, greatly can shorten sewage disposal start time, improves COD degradation efficiency.This domestication substratum has wider suitability, and after putting into multiple coal liquifaction sewage sample by the bacterial classification after its domestication, its COD degradation ability all increases.This test kit can simulate anoxic, aerobe method technique, can be applied to the products perfection of coal liquifaction sewage treatment microbial inoculant, and Sewage Plant is tamed fast to mud before operation.
The object of the invention is to be achieved through the following technical solutions: a kind of coal liquifaction waste water COD degraded strain domestication test kit, is characterized in that: it is made up of enrichment pipe, domestication prefabricated pipe A, domestication prefabricated pipe B; Wherein:
(1), enrichment pipe:
The preparation of enrichment pipe:
(1) extractum carnis 0.7-1.0g, is got, peptone 0.7-1.0g, potassium primary phosphate 1.2-2.0g, Sodium phosphate dibasic 1.5-2.5g, use 1L water dissolution, the sodium carbonate of above substratum concentration 5% and 5%HCl adjust pH to 7.00-7.50, and be dispensed in 50ml screw socket pipe, every arm adds substratum 20ml; After packing, unscrewed by screw socket pipe lid, 115 DEG C of sterilizing 25-30min, end subject to sterilization also, after being cooled to room temperature, being taken out and tightens pipe lid from Autoclave, and so far basal enrichment control is standby to be completed;
(2), get phenol 10-13mg, naphthols 2-4mg, quinoline 1-2mg, methyl alcohol 0.08-0.12ml, butanols 0.08-0.12ml, dissolve with 100ml sterilized water, after dissolving, aseptically use 0.22 μm of sterilised membrane filter filtration sterilization; After filtration sterilization completes, get the solution of 2ml through filtration sterilization, 0.2-0.3 μ l coal derived oil, aseptically join in the basal enrichment pipe in step (1), high speed whirlpool oscillator shakes 5-10min, and so far prepared by enrichment pipe;
(2), prefabricated pipe A is tamed:
The preparation of domestication prefabricated pipe A:
(1) extractum carnis 0.4-0.7g, is got, peptone 0.4-0.7g, glycerol 0.4-0.6ml, potassium primary phosphate 1.2-2.0g, Sodium phosphate dibasic 1.5-2.5g, uses 1L water dissolution, and the sodium carbonate of above substratum concentration 5% and 5%HCl adjust pH to 7.50-8.00, be dispensed in 50ml screw socket pipe, every arm adds substratum 30ml; After packing, unscrewed by screw socket pipe lid, 115 DEG C of sterilizing 25-30min, end subject to sterilization also, after being cooled to room temperature, being taken out and tightens pipe lid from Autoclave, prepared by so far basis domestication prefabricated pipe A;
(2) phenol 18-22mg, is got, naphthols 8-12mg, quinoline 4-6mg, aniline 4-6mg, pyridine 0.4-0.6mg, Standamul G 0.1-0.2mg, dissolve with 100ml sterilized water, aseptically use 0.22 μm of sterilised membrane filter filtration sterilization after dissolving, the solution after filtration sterilization is water additive A;
(3), get naphthalene 130-170mg, toluene 3-7mg, dibutyl phthalate 2-3mg, benzoglyoxaline 2-3mg, methyl alcohol 0.08-0.12ml, butanols 0.08-0.12ml, use 100ml75% dissolve with ethanol, this solution is alcohol additive A;
(4), fetch water additive A3ml respectively, alcohol additive A0.012ml, coal derived oil 1.0-1.2 μ l, aseptically installs in the basis domestication prefabricated pipe A in step (1), high speed whirlpool oscillator shakes 5-10min, so far tames prefabricated pipe A and prepared;
(3), prefabricated pipe B is tamed:
The preparation of domestication prefabricated pipe B:
(1) extractum carnis 0.2-0.4g, is got, peptone 0.2-0.4g, glycerol 0.6-0.8ml, potassium primary phosphate 1.2-2.0g, Sodium phosphate dibasic 1.5-2.5g, uses 1L water dissolution, and the sodium carbonate of above substratum concentration 5% and 5%HCl adjust pH to 7.50-8.00, be dispensed in 50ml screw socket pipe, every arm adds substratum 30ml; After packing, unscrewed by screw socket pipe lid, 115 DEG C of sterilizing 25-30min, end subject to sterilization also, after being cooled to room temperature, being taken out and tightens pipe lid from Autoclave, prepared by so far basis domestication prefabricated pipe B;
(2) phenol 150-170mg, is got, naphthols 18-22mg, quinoline 9-12mg, aniline 10-12mg, pyridine 0.9-1.2mg, Standamul G 0.4-0.6mg, dissolve with 100ml sterilized water, aseptically use 0.22 μm of sterilised membrane filter filtration sterilization after dissolving, the solution after filtration sterilization is water additive B;
(3), get naphthalene 540-560mg, toluene 18-22mg, dibutyl phthalate 8-12mg, benzoglyoxaline 8-12mg, methyl alcohol 1.8-2.2ml, butanols 1.8-2.2ml, use 100ml75% dissolve with ethanol, this solution is alcohol additive B;
(4), fetch water additive B3ml respectively, alcohol additive B0.012ml, coal derived oil 3.0-4.0 μ l, aseptically installs in the basis domestication prefabricated pipe B in step (1), high speed whirlpool oscillator shakes 5-10min, so far tames prefabricated pipe B and prepared.
A kind of coal liquifaction waste water COD that adopts is degraded with strain domestication test kit acclimation method, it is characterized in that: comprise the following steps:
(1), anoxic phases domestication:
Step 1, will treat that domestication sample is got 1-3ml and is inoculated in the 1st enrichment pipe, the pipe lid of the 1st enrichment pipe will be tightened, keep flat into shaking table 28-35 DEG C, 150-200rpm concussion cultivates 24-48 hour;
The sample 5ml that step 2, the 1st enrichment pipe of learning from else's experience are cultivated, is inoculated in the 1st domestication prefabricated pipe A, the 1st enrichment pipe is put into 4 DEG C of refrigerations together with remaining sample in pipe;
Step 3, by the 1st domestication prefabricated pipe A pipe lid tighten, keep flat into shaking table 25-30 DEG C, 100-150rpm concussion cultivate 24-48 hour; Standing 30-60min after cultivation completes, gets supernatant A 130ml and puts into 1st triangular flask of 100ml through sterilizing, then put into 4 DEG C of retentions; Get precipitation 5ml and be inoculated into continuation cultivation in the 2nd domestication prefabricated pipe A;
Step 4, by the 2nd domestication prefabricated pipe A pipe lid tighten, keep flat into shaking table 25-30 DEG C, 100-150rpm concussion cultivate 24-48 hour; Standing 30-60min after cultivation completes, gets supernatant A 230ml and puts into 2nd triangular flask of 100ml through sterilizing, then put into 4 DEG C of retentions; Get precipitation 5ml and be inoculated into continuation cultivation in the 3rd domestication prefabricated pipe A;
Step 5, by the 3rd domestication prefabricated pipe A pipe lid tighten, keep flat into shaking table 25-30 DEG C, 100-150rpm concussion cultivate 24-48 hour; Standing 30-60min after cultivation completes, gets supernatant A 330ml and puts into 3rd triangular flask of 100ml through sterilizing, then put into 4 DEG C of retentions; When sample is after the 3rd domestication prefabricated pipe A of step 4 cultivates, get precipitation 5ml, be inoculated in the 1st domestication prefabricated pipe B;
Step 6, by the 1st of step 5 the domestication prefabricated pipe B pipe lid tighten, keep flat into shaking table 25-28 DEG C, 100-150rpm concussion cultivate 48-72h; Standing 30-60min after cultivation completes, gets supernatant liquor B130ml and puts into 4th triangular flask of 100ml through sterilizing, then put into 4 DEG C of retentions; Get precipitation 5ml and be inoculated into continuation cultivation in the 2nd domestication prefabricated pipe B;
Step 7, by the 2nd of step 6 the domestication prefabricated pipe B pipe lid tighten, keep flat into shaking table 25-28 DEG C, 100-150rpm concussion cultivate 48-72h; Standing 30-60min after cultivation completes, gets supernatant liquor B230ml and puts into 5th triangular flask of 100ml through sterilizing, then put into 4 DEG C of retentions; Get precipitation 5ml and be inoculated into continuation cultivation in the 3rd domestication prefabricated pipe B;
Step 8, by the 3rd of step 7 the domestication prefabricated pipe B pipe lid tighten, keep flat into shaking table 25-28 DEG C, 100-150rpm concussion cultivate 48-72h; Standing 30-60min after cultivation completes, gets supernatant liquor B330ml and puts into 6th triangular flask of 100ml through sterilizing, then put into 4 DEG C of retentions; When sample is after the 3rd domestication prefabricated pipe B of step 7 cultivates, gets precipitation 1ml, be inoculated in the 2nd enrichment pipe, the pipe lid of the 2nd enrichment pipe is tightened, keep flat into shaking table 28-35 DEG C, 150-200rpm concussion cultivation 24-48 hour; By the 2nd enrichment pipe refrigeration or freezen protective after cultivation, after this domestication, bacterial classification is anoxic domesticated strain;
(2), aerobic stage domestication:
Step 9, after prefabricated pipe A cultivated through the 3rd domestication of step 5 when sample, the 1st enrichment pipe refrigerated is taken out and recovers room temperature; After room temperature 6-10h to be restored, get 5ml and be inoculated in supernatant A 3;
Step 10, postvaccinal for step 9 supernatant A 3 put into shaking table 25-28 DEG C, 150-200rpm concussion cultivates 24-48 hour; Standing 30-60min after cultivation completes, discards 30ml supernatant liquor, gets precipitation 5ml and is inoculated into continuation cultivation in supernatant A 2;
Step 11, postvaccinal for step 10 supernatant A 2 put into shaking table 25-28 DEG C, 150-200rpm concussion cultivates 24-48 hour; Standing 30-60min after cultivation completes, discards 30ml supernatant liquor, gets precipitation 5ml and is inoculated into continuation cultivation in supernatant A 1;
Step 12, postvaccinal for step 11 supernatant A 1 put into shaking table 25-28 DEG C, 150-200rpm concussion cultivates 24-48 hour; Standing 30-60min after cultivation completes, discards 30ml supernatant liquor, when sample is tamed after cultivation through supernatant A 1, gets precipitation 5ml, is inoculated in supernatant liquor B3;
Step 13, postvaccinal for step 12 supernatant liquor B3 entered shaking table 25-28 DEG C, 150-200rpm concussion cultivates 48-72 hour; Cultivation leaves standstill after completing
-60min, discards 30ml supernatant liquor, gets precipitation 5ml and is inoculated into continuation cultivation in supernatant liquor B2;
Step 14, postvaccinal for step 13 supernatant liquor B2 entered shaking table 25-28 DEG C, 150-200rpm concussion cultivates 48-72 hour; Standing 30-60min after cultivation completes, discards 30ml supernatant liquor, gets precipitation 5ml and is inoculated into continuation cultivation in supernatant liquor B1;
Step 15, postvaccinal for step 14 supernatant liquor B1 entered shaking table 25-28 DEG C, 150-200rpm concussion cultivates 48-72 hour; Standing 30-60min after cultivation completes, discards 30ml supernatant liquor, when sample is tamed after cultivation through step supernatant liquor B1, get precipitation 1ml, be inoculated in the 3rd enrichment pipe, the pipe lid of the 3rd enrichment pipe is tightened, keep flat into shaking table 28-35 DEG C, 150-200rpm concussion cultivation 24-48 hour; By the 3rd enrichment pipe refrigeration or freezen protective after cultivation, after this domestication, bacterial classification is aerobic domesticated strain.
Advantage of the present invention and beneficial effect are:
This test kit gathers coal liquifaction wastewater sample by this laboratory in a large number in the whole nation, waste component is analyzed, and make by experiment a kind of there is broad applicability can the coal liquifaction waste water strain domestication substratum of Reality simulation sewage, and define test kit based on this, through the bacterial classification of its domestication, can as efficient coal liquifaction waste water COD process microbial inoculum or mud, improve the adaptive faculty of bacterial classification in waste water, shorten sewage disposal start time.
Accompanying drawing illustrates:
Fig. 1 is acclimation method schema of the present invention.
Wherein:
1-sample, 2-the 1st enrichment pipe, 3-the 1st tames prefabricated pipe A, 4-the 2nd domestication prefabricated pipe A, 5-the 3rd tames prefabricated pipe A, 6-the 1st and tames prefabricated pipe B, 7-the 2nd tames prefabricated pipe B, 8-the 3rd domestication prefabricated pipe B, 9-the 1st triangular flask, 10-the 2nd triangular flask, 11-the 3rd triangular flask, 12-the 4th triangular flask, 13-the 5th triangular flask, 14-the 6th triangular flask, 15-the 2nd enrichment pipe, 16-the 3rd enrichment pipe.
Embodiment:
Below in conjunction with specific embodiment, set forth the present invention further.These embodiments should be understood only be not used in for illustration of the present invention and limit the scope of the invention.It should be noted that: the sterilizing of operating environment and relevant appliance before operation, should be guaranteed.Raw material of the present invention is commercially available prod if no special instructions.Particularly point out, illustrate for 50ml prefabricated pipe specification below, but be not limited to 50ml specification.
Embodiment 1:
Embodiment 1 used kit formula and preparation method as follows:
One, enrichment pipe:
The preparation of enrichment pipe:
1, get extractum carnis 1.0g, peptone 1.0g, potassium primary phosphate 2.0g, Sodium phosphate dibasic 2.5g, use 1L water dissolution, the sodium carbonate of above substratum concentration 5% and 5%HCl adjust pH to 7.00, and be dispensed in 50ml screw socket pipe, every arm adds substratum 20ml; After packing, unscrewed by screw socket pipe lid, 115 DEG C of sterilizing 25min, end subject to sterilization also, after being cooled to room temperature, being taken out and tightens pipe lid from Autoclave, and so far basal enrichment control is standby to be completed;
2, get phenol 13mg, naphthols 4mg, quinoline 2mg, methyl alcohol 0.12ml, butanols 0.12ml, dissolve with 100ml sterilized water, after dissolving, aseptically use 0.22 μm of sterilised membrane filter filtration sterilization; After filtration sterilization completes, get the solution of 2ml through filtration sterilization, 0.2 μ l coal derived oil, aseptically join in the basal enrichment pipe in step 1, high speed whirlpool oscillator shakes 5min, and so far prepared by enrichment pipe;
Two, prefabricated pipe A is tamed:
The preparation of domestication prefabricated pipe A:
1, extractum carnis 0.7g is got, peptone 0.7g, glycerol 0.6ml, potassium primary phosphate 2.0g, Sodium phosphate dibasic 2.5g, use 1L water dissolution, the sodium carbonate of above substratum concentration 5% and 5%HCl adjust pH to 7.50, and be dispensed in 50ml screw socket pipe, every arm adds substratum 30ml; After packing, unscrewed by screw socket pipe lid, 115 DEG C of sterilizing 25min, end subject to sterilization also, after being cooled to room temperature, being taken out and tightens pipe lid from Autoclave, prepared by so far basis domestication prefabricated pipe A;
2, get phenol 22mg, naphthols 12mg, quinoline 6mg, aniline 6mg, pyridine 0.6mg, Standamul G 0.2mg, dissolve with 100ml sterilized water, aseptically use 0.22 μm of sterilised membrane filter filtration sterilization after dissolving, the solution after filtration sterilization is water additive A;
3, get naphthalene 170mg, toluene 7mg, dibutyl phthalate 3mg, benzoglyoxaline 3mg, methyl alcohol 0.12ml, butanols 0.12ml, use 100ml75% dissolve with ethanol, this solution is alcohol additive A;
4, fetch water additive A3ml, alcohol additive A0.012ml, coal derived oil 1.0 μ l respectively, aseptically installs in the basis domestication prefabricated pipe A in step 1, high speed whirlpool oscillator shakes 5min, so far tames prefabricated pipe A and prepared;
Three, prefabricated pipe B is tamed:
The preparation of domestication prefabricated pipe B:
1, extractum carnis 0.4g is got, peptone 0.4g, glycerol 0.8ml, potassium primary phosphate 2.0g, Sodium phosphate dibasic 2.5g, use 1L water dissolution, the sodium carbonate of above substratum concentration 5% and 5%HCl adjust pH to 7.50, and be dispensed in 50ml screw socket pipe, every arm adds substratum 30ml; After packing, unscrewed by screw socket pipe lid, 115 DEG C of sterilizing 25min, end subject to sterilization also, after being cooled to room temperature, being taken out and tightens pipe lid from Autoclave, prepared by so far basis domestication prefabricated pipe B;
2, phenol 170mg is got, naphthols 22mg, quinoline 12mg, aniline 12mg, pyridine 1.2mg, Standamul G 0.6mg, dissolve with 100ml sterilized water, aseptically use 0.22 μm of sterilised membrane filter filtration sterilization after dissolving, the solution after filtration sterilization is water additive B;
3, get naphthalene 560mg, toluene 22mg, dibutyl phthalate 12mg, benzoglyoxaline 12mg, methyl alcohol 2.2ml, butanols 2.2ml, use 100ml75% dissolve with ethanol, this solution is alcohol additive B;
4, fetch water additive B3ml, alcohol additive B0.012ml, coal derived oil 3.0 μ l respectively, aseptically installs in the basis domestication prefabricated pipe B in step 1, high speed whirlpool oscillator shakes 5min, so far tames prefabricated pipe B and prepared.
Adopt this test kit to the method for carrying out strain domestication, see Fig. 1.Wherein sample source is the sewage disposal liquid bacterial agent for the treatment of organic chemical waste water of this laboratory development, and microbial inoculum mainly forms and comprises: Rhodopseudomonas, acinetobacter, genus arthrobacter, gemma Pseudomonas etc.Detailed acclimation method is as follows:
One, anoxic phases domestication:
Step 1, will treat that domestication sample 1 is got 1ml and is inoculated in the 1st enrichment pipe 2, and the pipe lid of the 1st enrichment pipe 2 be tightened, keeps flat and shake cultivation 24 hours into shaking table 35 DEG C, 200rpm;
The sample 5ml of the 1st enrichment pipe 2 cultivation of step 2, step 1 of learning from else's experience, is inoculated in the 1st domestication prefabricated pipe A3, and the 1st enrichment pipe 2 is put into 4 DEG C of refrigerations together with remaining sample in pipe;
Step 3, the pipe lid of the 1st of step 2 the domestication prefabricated pipe A3 to be tightened, keep flat and shake cultivation 24 hours into shaking table 28 DEG C, 150rpm; Standing 60min after cultivation completes, gets supernatant liquor 30ml and puts into 1st triangular flask 9 of 100ml through sterilizing, then put into 4 DEG C of retentions; Get precipitation 5ml and be inoculated into continuation cultivation in the 2nd domestication prefabricated pipe A4;
Step 4, the pipe lid of the 2nd of step 3 the domestication prefabricated pipe A4 to be tightened, keep flat and shake cultivation 24 hours into shaking table 28 DEG C, 150rpm; Standing 60min after cultivation completes, gets supernatant liquor 30ml and puts into 2nd triangular flask 10 of 100ml through sterilizing, then put into 4 DEG C of retentions; Get precipitation 5ml and be inoculated into continuation cultivation in the 3rd domestication prefabricated pipe A5;
Step 5, the pipe lid of the 3rd of step 4 the domestication prefabricated pipe A5 to be tightened, keep flat and shake cultivation 24 hours into shaking table 28 DEG C, 150rpm; Standing 60min after cultivation completes, gets supernatant liquor 30ml and puts into 3rd triangular flask 11 of 100ml through sterilizing, then put into 4 DEG C of retentions; When sample is after the 3rd domestication prefabricated pipe A5 cultivates, get precipitation 5ml, be inoculated in the 1st domestication prefabricated pipe B6;
Step 6, by the 1st of step 5 the domestication prefabricated pipe B6 pipe lid tighten, keep flat into shaking table 28 DEG C, 150rpm concussion cultivate 48h; Standing 60min after cultivation completes, gets supernatant liquor 30ml and puts into 4th triangular flask 12 of 100ml through sterilizing, then put into 4 DEG C of retentions; Get precipitation 5ml and be inoculated into continuation cultivation in the 2nd domestication prefabricated pipe B7;
Step 7, by the 2nd of step 6 the domestication prefabricated pipe B7 pipe lid tighten, keep flat into shaking table 28 DEG C, 150rpm concussion cultivate 48h; Standing 60min after cultivation completes, gets supernatant liquor 30ml and puts into 5th triangular flask 13 of 100ml through sterilizing, then put into 4 DEG C of retentions; Get precipitation 5ml and be inoculated into continuation cultivation in the 3rd domestication prefabricated pipe B8;
Step 8, by the 3rd of step 7 the domestication prefabricated pipe B8 pipe lid tighten, keep flat into shaking table 28 DEG C, 150rpm concussion cultivate 48h; Standing 60min after cultivation completes, gets supernatant liquor 30ml and puts into 6th triangular flask 14 of 100ml through sterilizing, then put into 4 DEG C of retentions; When sample is after the 3rd domestication prefabricated pipe B8 cultivates, gets precipitation 1ml, be inoculated in the 2nd enrichment pipe 15, the pipe lid of the 2nd enrichment pipe 15 is tightened, keep flat and shake cultivation 24 hours into shaking table 35 DEG C, 200rpm; Cultivate in backward 2nd enrichment pipe 15 and add 4ml sterile glycerol, be dispensed into cryopreservation tube frozen after mixing, after this domestication, bacterial classification is anoxic domesticated strain;
Two, aerobic stage domestication:
Step 9, after prefabricated pipe A5 cultivated through the 3rd domestication of step 5 when sample, the 1st enrichment pipe 2 refrigerated is taken out and recovers room temperature; After room temperature 6h to be restored, get 5ml and be inoculated in supernatant A 3;
Step 10, postvaccinal for step 9 supernatant A 3 entered shaking table 28 DEG C, 200rpm shakes cultivation 24 hours; Standing 60min after cultivation completes, discards 30ml supernatant liquor, gets precipitation 5ml and is inoculated into continuation cultivation in supernatant A 2;
Step 11, postvaccinal for step 10 supernatant A 2 entered shaking table 28 DEG C, 200rpm shakes cultivation 24 hours; Standing 60min after cultivation completes, discards 30ml supernatant liquor, gets precipitation 5ml and is inoculated into continuation cultivation in supernatant A 1;
Step 12, postvaccinal for step 11 supernatant A 1 entered shaking table 28 DEG C, 200rpm shakes cultivation 24 hours; Standing 60min after cultivation completes, discards 30ml supernatant liquor, when sample is after the supernatant A 1 of step 11 tames cultivation, gets precipitation 5ml, is inoculated in supernatant liquor B3;
Step 13, postvaccinal for step 12 supernatant liquor B3 put into shaking table 28 DEG C, 200rpm shakes cultivation 48 hours; Standing 60min after cultivation completes, discards 30ml supernatant liquor, gets precipitation 5ml and is inoculated into continuation cultivation in supernatant liquor B2;
Step 14, postvaccinal for step 13 supernatant liquor B2 put into shaking table 28 DEG C, 200rpm shakes cultivation 48 hours; Standing 60min after cultivation completes, discards 30ml supernatant liquor, gets precipitation 5ml and is inoculated into continuation cultivation in supernatant liquor B1;
Step 15, postvaccinal for step 14 supernatant liquor B1 put into shaking table 28 DEG C, 200rpm shakes cultivation 48 hours; Standing 60min after cultivation completes, discards 30ml supernatant liquor, when sample is tamed after cultivation through supernatant liquor B1, gets precipitation 1ml, is inoculated in the 3rd enrichment pipe 16, is tightened by the pipe lid of the 3rd enrichment pipe 16, keep flat and shake cultivation 24 hours into shaking table 35 DEG C, 200rpm; Cultivate in backward 3rd enrichment pipe and add 4ml sterile glycerol, frozen after being dispensed into cryopreservation tube after mixing, after this domestication, bacterial classification is aerobic domesticated strain.
Bacterial classification through domestication carries out domestication effect comparison by following experiment:
Sewage source is the equalizing tank water outlet of the Inner Mongol one coal liquifaction enterprise, and COD is 3948mg/L.Respectively getting sewage 40ml is distributed in 2 50ml aseptic screw socket pipes stand-by.Sample to be verified is through the bacterial classification of domestication and the bacterial classification without domestication.Verification method is: 1, anoxic compliance test result: by the bacterial classification without domestication and the bacterial classification through anoxic domestication by 2% inoculum size be inoculated into respectively and be equipped with in the screw socket pipe of sewage, keep flat and shake cultivation into shaking table 28 DEG C, 100rpm, survey a COD every day, Continuous Observation 5 days.2, aerobic compliance test result: the above sewage through anoxic checking is proceeded in 2 sterilized triangular flasks respectively, by without domestication bacterial classification and through aerobic domestication bacterial classification by 2% inoculum size be inoculated in 2 triangular flasks respectively, put into shaking table 28 DEG C, 200rpm shakes cultivation, survey a COD every day, Continuous Observation 5 days.Observations is as follows.
Table 1: bacterial classification effect comparison before and after domestication
Can be found out by simultaneous test, the bacterial classification through test kit domestication has toggle speed and better degradation efficiency faster.
Embodiment 2:
Embodiment 2 used kit formula and preparation method as follows:
One, enrichment pipe:
The preparation of enrichment pipe:
1, get extractum carnis 0.7g, peptone 0.7g, potassium primary phosphate 1.2g, Sodium phosphate dibasic 1.5g, use 1L water dissolution, the sodium carbonate of above substratum concentration 5% and 5%HCl adjust pH to 7.50, and be dispensed in 50ml screw socket pipe, every arm adds substratum 20ml; After packing, unscrewed by screw socket pipe lid, 115 DEG C of sterilizing 30min, end subject to sterilization also, after being cooled to room temperature, being taken out and tightens pipe lid from Autoclave, and so far basal enrichment control is standby to be completed;
2, get phenol 10mg, naphthols 2mg, quinoline 1mg, methyl alcohol 0.08ml, butanols 0.08ml, dissolve with 100ml sterilized water, after dissolving, aseptically use 0.22 μm of sterilised membrane filter filtration sterilization; After filtration sterilization completes, get the solution of 2ml through filtration sterilization, 0.3 μ l coal derived oil, aseptically join in the basal enrichment pipe in step 1, high speed whirlpool oscillator shakes 10min, and so far prepared by enrichment pipe;
Two, prefabricated pipe A is tamed:
The preparation of domestication prefabricated pipe A:
1, extractum carnis 0.4g is got, peptone 0.4g, glycerol 0.4ml, potassium primary phosphate 1.2g, Sodium phosphate dibasic 1.5g, use 1L water dissolution, the sodium carbonate of above substratum concentration 5% and 5%HCl adjust pH to 8.00, and be dispensed in 50ml screw socket pipe, every arm adds substratum 30ml; After packing, unscrewed by screw socket pipe lid, 115 DEG C of sterilizing 30min, end subject to sterilization also, after being cooled to room temperature, being taken out and tightens pipe lid from Autoclave, prepared by so far basis domestication prefabricated pipe A;
2, get phenol 18mg, naphthols 8mg, quinoline 4mg, aniline 4mg, pyridine 0.4mg, Standamul G 0.1mg, dissolve with 100ml sterilized water, aseptically use 0.22 μm of sterilised membrane filter filtration sterilization after dissolving, the solution after filtration sterilization is water additive A;
3, get naphthalene 130mg, toluene 3mg, dibutyl phthalate 2mg, benzoglyoxaline 2mg, methyl alcohol 0.08ml, butanols 0.08ml, use 100ml75% dissolve with ethanol, this solution is alcohol additive A;
4, fetch water additive A3ml, alcohol additive A0.012ml, coal derived oil 1.2 μ l respectively, aseptically installs in the basis domestication prefabricated pipe A in step 1, high speed whirlpool oscillator shakes 10min, so far tames prefabricated pipe A and prepared;
Three, prefabricated pipe B is tamed:
The preparation of domestication prefabricated pipe B:
1, extractum carnis 0.2g is got, peptone 0.2g, glycerol 0.6ml, potassium primary phosphate 1.2g, Sodium phosphate dibasic 1.5g, use 1L water dissolution, the sodium carbonate of above substratum concentration 5% and 5%HCl adjust pH to 8.00, and be dispensed in 50ml screw socket pipe, every arm adds substratum 30ml; After packing, unscrewed by screw socket pipe lid, 115 DEG C of sterilizing 30min, end subject to sterilization also, after being cooled to room temperature, being taken out and tightens pipe lid from Autoclave, prepared by so far basis domestication prefabricated pipe B;
2, phenol 150mg is got, naphthols 18mg, quinoline 9mg, aniline 10mg, pyridine 0.9mg, Standamul G 0.4mg, dissolve with 100ml sterilized water, aseptically use 0.22 μm of sterilised membrane filter filtration sterilization after dissolving, the solution after filtration sterilization is water additive B;
3, get naphthalene 540mg, toluene 18mg, dibutyl phthalate 8mg, benzoglyoxaline 8mg, methyl alcohol 1.8ml, butanols 1.8ml, use 100ml75% dissolve with ethanol, this solution is alcohol additive B;
4, fetch water additive B3ml, alcohol additive B0.012ml, coal derived oil 4.0 μ l respectively, aseptically installs in the basis domestication prefabricated pipe B in step 1, high speed whirlpool oscillator shakes 10min, so far tames prefabricated pipe B and prepared.
This test kit is adopted to see Fig. 1 to the method that bacterial classification is tamed.Wherein sample source is the activated sludge sample that this laboratory gathers from chemical engineering industry garden, Shandong one sewage work.Method detailed is as follows:
One, anoxic phases domestication:
Step 1, will treat that domestication sample 1 is got 3ml and is inoculated in the 1st enrichment pipe 2, and the pipe lid of the 1st enrichment pipe 2 be tightened, keeps flat and shake cultivation 48 hours into shaking table 28 DEG C, 150rpm;
The sample 5ml of the 1st enrichment pipe 2 cultivation of step 2, step 1 of learning from else's experience, is inoculated in the 1st domestication prefabricated pipe A3, the 1st enrichment pipe 2 is put into 4 DEG C of refrigerations together with remaining sample in pipe;
Step 3, the pipe lid of the 1st of step 2 the domestication prefabricated pipe A3 to be tightened, keep flat and shake cultivation 48 hours into shaking table 25 DEG C, 100rpm; Standing 30min after cultivation completes, gets supernatant liquor 30ml and puts into 1st triangular flask 9 of 100ml through sterilizing, then put into 4 DEG C of retentions; Get precipitation 5ml and be inoculated into continuation cultivation in the 2nd domestication prefabricated pipe A4;
Step 4, the pipe lid of the 2nd of step 3 the domestication prefabricated pipe A4 to be tightened, keep flat and shake cultivation 48 hours into shaking table 25 DEG C, 100rpm; Standing 30min after cultivation completes, gets supernatant liquor 30ml and puts into 2nd triangular flask 10 of 100ml through sterilizing, then put into 4 DEG C of retentions; Get precipitation 5ml and be inoculated into continuation cultivation in the 3rd domestication prefabricated pipe A5;
Step 5, the pipe lid of the 3rd of step 4 the domestication prefabricated pipe A5 to be tightened, keep flat and shake cultivation 48 hours into shaking table 25 DEG C, 100rpm; Standing 30min after cultivation completes, gets supernatant liquor 30ml and puts into 3rd triangular flask 11 of 100ml through sterilizing, then put into 4 DEG C of retentions; When sample is after the 3rd domestication prefabricated pipe A5 cultivates, get precipitation 5ml, be inoculated in the 1st domestication prefabricated pipe B6;
Step 6, by the 1st of step 5 the domestication prefabricated pipe B6 pipe lid tighten, keep flat into shaking table 25 DEG C, 100rpm concussion cultivate 72h; Standing 30min after cultivation completes, gets supernatant liquor 30ml and puts into 4th triangular flask 12 of 100ml through sterilizing, then put into 4 DEG C of retentions; Get precipitation 5ml and be inoculated into continuation cultivation in the 2nd domestication prefabricated pipe B7;
Step 7, by the 2nd of step 6 the domestication prefabricated pipe B7 pipe lid tighten, keep flat into shaking table 25 DEG C, 100rpm concussion cultivate 72h; Standing 30min after cultivation completes, gets supernatant liquor 30ml and puts into 5th triangular flask 13 of 100ml through sterilizing, then put into 4 DEG C of retentions; Get precipitation 5ml and be inoculated into continuation cultivation in the 3rd domestication prefabricated pipe B8;
Step 8, by the 3rd of step 7 the domestication prefabricated pipe B8 pipe lid tighten, keep flat into shaking table 25 DEG C, 100rpm concussion cultivate 72h; Standing 30min after cultivation completes, gets supernatant liquor 30ml and puts into 6th triangular flask 14 of 100ml through sterilizing, then put into 4 DEG C of retentions; When sample is after the 3rd domestication prefabricated pipe B8 cultivates, gets precipitation 1ml, be inoculated in the 2nd enrichment pipe 15, the pipe lid of the 2nd enrichment pipe 15 is tightened, keep flat and shake cultivation 48 hours into shaking table 28 DEG C, 150rpm; After cultivation, the 2nd enrichment pipe 15 is put into 4 DEG C of preservations, after this domestication, bacterial classification is anoxic domesticated strain;
Two, aerobic stage domestication:
Step 9, after prefabricated pipe A5 cultivated through the 3rd domestication of step 5 when sample, the 1st enrichment pipe 2 refrigerated is taken out and recovers room temperature; After room temperature 10h to be restored, get 5ml and be inoculated in supernatant A 3;
Step 10, postvaccinal for step 9 supernatant A 3 entered shaking table 25 DEG C, 150rpm shakes cultivation 48 hours; Standing 30min after cultivation completes, discards 30ml supernatant liquor, gets precipitation 5ml and is inoculated into continuation cultivation in supernatant A 2;
Step 11, postvaccinal for step 10 supernatant A 2 entered shaking table 25 DEG C, 150rpm shakes cultivation 48 hours; Standing 30min after cultivation completes, discards 30ml supernatant liquor, gets precipitation 5ml and is inoculated into continuation cultivation in supernatant A 1;
Step 12, postvaccinal for step 11 supernatant A 1 entered shaking table 25 DEG C, 150rpm shakes cultivation 48 hours; Standing 30min after cultivation completes, discards 30ml supernatant liquor, when sample is tamed after cultivation through supernatant A 1, gets precipitation 5ml, is inoculated in supernatant liquor B3;
Step 13, postvaccinal for step 12 supernatant liquor B3 put into shaking table 25 DEG C, 150rpm shakes cultivation 72 hours; Standing 30min after cultivation completes, discards 30ml supernatant liquor, gets precipitation 5ml and is inoculated into continuation cultivation in supernatant liquor B2;
Step 14, postvaccinal for step 13 supernatant liquor B2 put into shaking table 25 DEG C, 150rpm shakes cultivation 72 hours; Standing 30min after cultivation completes, discards 30ml supernatant liquor, gets precipitation 5ml and is inoculated into continuation cultivation in supernatant liquor B1;
Step 15, postvaccinal for step 14 supernatant liquor B1 put into shaking table 25 DEG C, 150rpm shakes cultivation 72 hours; Standing 30min after cultivation completes, discards 30ml supernatant liquor, when sample is tamed after cultivation through supernatant liquor B1, gets precipitation 1ml, is inoculated in the 3rd enrichment pipe 16, is tightened by the pipe lid of the 3rd enrichment pipe 16, keep flat and shake cultivation 48 hours into shaking table 28 DEG C, 150rpm; After cultivation, the 3rd enrichment pipe 16 is put into 4 DEG C of preservations, after this domestication, bacterial classification is aerobic domesticated strain.
Bacterial classification through domestication carries out domestication effect comparison by following experiment:
Sewage source is the equalizing tank water outlet of the Inner Mongol one coal liquifaction enterprise, and COD is 3782mg/L.Respectively getting sewage 40ml is distributed in 2 50ml aseptic screw socket pipes stand-by.Sample to be verified is through the bacterial classification of domestication and the bacterial classification without domestication.Verification method is: 1, anoxic compliance test result: by the bacterial classification without domestication and the bacterial classification through anoxic domestication by 2% inoculum size be inoculated into respectively and be equipped with in the screw socket pipe of sewage, keep flat and shake cultivation into shaking table 28 DEG C, 100rpm, survey a COD every day, Continuous Observation 5 days.2, aerobic compliance test result: the above sewage through anoxic checking is proceeded in 2 sterilized triangular flasks respectively, by without domestication bacterial classification and through aerobic domestication bacterial classification by 2% inoculum size be inoculated in 2 triangular flasks respectively, put into shaking table 28 DEG C, 200rpm shakes cultivation, survey a COD every day, Continuous Observation 5 days.Observations is as follows.
Table 2: bacterial classification effect comparison before and after domestication
Can be found out by simultaneous test, the bacterial classification through test kit domestication has toggle speed and better degradation efficiency faster.
Claims (2)
1. a strain domestication test kit is used in the degraded of coal liquifaction waste water COD, it is characterized in that: it is made up of enrichment pipe, domestication prefabricated pipe A, domestication prefabricated pipe B; Wherein:
(1), enrichment pipe:
The preparation of enrichment pipe:
(1) extractum carnis 0.7-1.0g, is got, peptone 0.7-1.0g, potassium primary phosphate 1.2-2.0g, Sodium phosphate dibasic 1.5-2.5g, use 1L water dissolution, the sodium carbonate of above substratum concentration 5% and 5%HCl adjust pH to 7.00-7.50, and be dispensed in 50ml screw socket pipe, every arm adds substratum 20ml; After packing, unscrewed by screw socket pipe lid, 115 DEG C of sterilizing 25-30min, end subject to sterilization also, after being cooled to room temperature, being taken out and tightens pipe lid from Autoclave, and so far basal enrichment control is standby to be completed;
(2), get phenol 10-13mg, naphthols 2-4mg, quinoline 1-2mg, methyl alcohol 0.08-0.12ml, butanols 0.08-0.12ml, dissolve with 100ml sterilized water, after dissolving, aseptically use 0.22 μm of sterilised membrane filter filtration sterilization; After filtration sterilization completes, get the solution of 2ml through filtration sterilization, 0.2-0.3 μ l coal derived oil, aseptically join in the basal enrichment pipe in step (1), high speed whirlpool oscillator shakes 5-10min, and so far prepared by enrichment pipe;
(2), prefabricated pipe A is tamed:
The preparation of domestication prefabricated pipe A:
(1) extractum carnis 0.4-0.7g, is got, peptone 0.4-0.7g, glycerol 0.4-0.6ml, potassium primary phosphate 1.2-2.0g, Sodium phosphate dibasic 1.5-2.5g, uses 1L water dissolution, and the sodium carbonate of above substratum concentration 5% and 5%HCl adjust pH to 7.50-8.00, be dispensed in 50ml screw socket pipe, every arm adds substratum 30ml; After packing, unscrewed by screw socket pipe lid, 115 DEG C of sterilizing 25-30min, end subject to sterilization also, after being cooled to room temperature, being taken out and tightens pipe lid from Autoclave, prepared by so far basis domestication prefabricated pipe A;
(2) phenol 18-22mg, is got, naphthols 8-12mg, quinoline 4-6mg, aniline 4-6mg, pyridine 0.4-0.6mg, Standamul G 0.1-0.2mg, dissolve with 100ml sterilized water, aseptically use 0.22 μm of sterilised membrane filter filtration sterilization after dissolving, the solution after filtration sterilization is water additive A;
(3), get naphthalene 130-170mg, toluene 3-7mg, dibutyl phthalate 2-3mg, benzoglyoxaline 2-3mg, methyl alcohol 0.08-0.12ml, butanols 0.08-0.12ml, use 100ml75% dissolve with ethanol, this solution is alcohol additive A;
(4), fetch water additive A3ml respectively, alcohol additive A0.012ml, coal derived oil 1.0-1.2 μ l, aseptically installs in the basis domestication prefabricated pipe A in step (1), high speed whirlpool oscillator shakes 5-10min, so far tames prefabricated pipe A and prepared;
(3), prefabricated pipe B is tamed:
The preparation of domestication prefabricated pipe B:
(1) extractum carnis 0.2-0.4g, is got, peptone 0.2-0.4g, glycerol 0.6-0.8ml, potassium primary phosphate 1.2-2.0g, Sodium phosphate dibasic 1.5-2.5g, uses 1L water dissolution, and the sodium carbonate of above substratum concentration 5% and 5%HCl adjust pH to 7.50-8.00, be dispensed in 50ml screw socket pipe, every arm adds substratum 30ml; After packing, unscrewed by screw socket pipe lid, 115 DEG C of sterilizing 25-30min, end subject to sterilization also, after being cooled to room temperature, being taken out and tightens pipe lid from Autoclave, prepared by so far basis domestication prefabricated pipe B;
(2) phenol 150-170mg, is got, naphthols 18-22mg, quinoline 9-12mg, aniline 10-12mg, pyridine 0.9-1.2mg, Standamul G 0.4-0.6mg, dissolve with 100ml sterilized water, aseptically use 0.22 μm of sterilised membrane filter filtration sterilization after dissolving, the solution after filtration sterilization is water additive B;
(3), get naphthalene 540-560mg, toluene 18-22mg, dibutyl phthalate 8-12mg, benzoglyoxaline 8-12mg, methyl alcohol 1.8-2.2ml, butanols 1.8-2.2ml, use 100ml75% dissolve with ethanol, this solution is alcohol additive B;
(4), fetch water additive B3ml respectively, alcohol additive B0.012ml, coal derived oil 3.0-4.0 μ l, aseptically installs in the basis domestication prefabricated pipe B in step (1), high speed whirlpool oscillator shakes 5-10min, so far tames prefabricated pipe B and prepared.
2. adopt a coal liquifaction waste water COD degraded strain domestication test kit acclimation method according to claim 1, it is characterized in that: comprise the following steps:
(1), anoxic phases domestication:
Step 1, will treat that domestication sample is got 1-3ml and is inoculated in the 1st enrichment pipe, the pipe lid of the 1st enrichment pipe will be tightened, keep flat into shaking table 28-35 DEG C, 150-200rpm concussion cultivates 24-48 hour;
The sample 5ml that step 2, the 1st enrichment pipe of learning from else's experience are cultivated, is inoculated in the 1st domestication prefabricated pipe A, the 1st enrichment pipe is put into 4 DEG C of refrigerations together with remaining sample in pipe;
Step 3, by the 1st domestication prefabricated pipe A pipe lid tighten, keep flat into shaking table 25-30 DEG C, 100-150rpm concussion cultivate 24-48 hour; Standing 30-60min after cultivation completes, gets supernatant A 130ml and puts into 1st triangular flask of 100ml through sterilizing, then put into 4 DEG C of retentions; Get precipitation 5ml and be inoculated into continuation cultivation in the 2nd domestication prefabricated pipe A;
Step 4, by the 2nd domestication prefabricated pipe A pipe lid tighten, keep flat into shaking table 25-30 DEG C, 100-150rpm concussion cultivate 24-48 hour; Standing 30-60min after cultivation completes, gets supernatant A 230ml and puts into 2nd triangular flask of 100ml through sterilizing, then put into 4 DEG C of retentions; Get precipitation 5ml and be inoculated into continuation cultivation in the 3rd domestication prefabricated pipe A;
Step 5, by the 3rd domestication prefabricated pipe A pipe lid tighten, keep flat into shaking table 25-30 DEG C, 100-150rpm concussion cultivate 24-48 hour; Standing 30-60min after cultivation completes, gets supernatant A 330ml and puts into 3rd triangular flask of 100ml through sterilizing, then put into 4 DEG C of retentions; When sample is after the 3rd domestication prefabricated pipe A of step 4 cultivates, get precipitation 5ml, be inoculated in the 1st domestication prefabricated pipe B;
Step 6, by the 1st of step 5 the domestication prefabricated pipe B pipe lid tighten, keep flat into shaking table 25-28 DEG C, 100-150rpm concussion cultivate 48-72h; Standing 30-60min after cultivation completes, gets supernatant liquor B130ml and puts into 4th triangular flask of 100ml through sterilizing, then put into 4 DEG C of retentions; Get precipitation 5ml and be inoculated into continuation cultivation in the 2nd domestication prefabricated pipe B;
Step 7, by the 2nd of step 6 the domestication prefabricated pipe B pipe lid tighten, keep flat into shaking table 25-28 DEG C, 100-150rpm concussion cultivate 48-72h; Standing 30-60min after cultivation completes, gets supernatant liquor B230ml and puts into 5th triangular flask of 100ml through sterilizing, then put into 4 DEG C of retentions; Get precipitation 5ml and be inoculated into continuation cultivation in the 3rd domestication prefabricated pipe B;
Step 8, by the 3rd of step 7 the domestication prefabricated pipe B pipe lid tighten, keep flat into shaking table 25-28 DEG C, 100-150rpm concussion cultivate 48-72h; Standing 30-60min after cultivation completes, gets supernatant liquor B330ml and puts into 6th triangular flask of 100ml through sterilizing, then put into 4 DEG C of retentions; When sample is after the 3rd domestication prefabricated pipe B of step 7 cultivates, gets precipitation 1ml, be inoculated in the 2nd enrichment pipe, the pipe lid of the 2nd enrichment pipe is tightened, keep flat into shaking table 28-35 DEG C, 150-200rpm concussion cultivation 24-48 hour; By the 2nd enrichment pipe refrigeration or freezen protective after cultivation, after this domestication, bacterial classification is anoxic domesticated strain;
(2), aerobic stage domestication:
Step 9, after prefabricated pipe A cultivated through the 3rd domestication of step 5 when sample, the 1st enrichment pipe refrigerated is taken out and recovers room temperature; After room temperature 6-10h to be restored, get 5ml and be inoculated in supernatant A 3;
Step 10, postvaccinal for step 9 supernatant A 3 put into shaking table 25-28 DEG C, 150-200rpm concussion cultivates 24-48 hour; Standing 30-60min after cultivation completes, discards 30ml supernatant liquor, gets precipitation 5ml and is inoculated into continuation cultivation in supernatant A 2;
Step 11, postvaccinal for step 10 supernatant A 2 put into shaking table 25-28 DEG C, 150-200rpm concussion cultivates 24-48 hour; Standing 30-60min after cultivation completes, discards 30ml supernatant liquor, gets precipitation 5ml and is inoculated into continuation cultivation in supernatant A 1;
Step 12, postvaccinal for step 11 supernatant A 1 put into shaking table 25-28 DEG C, 150-200rpm concussion cultivates 24-48 hour; Standing 30-60min after cultivation completes, discards 30ml supernatant liquor, when sample is tamed after cultivation through supernatant A 1, gets precipitation 5ml, is inoculated in supernatant liquor B3;
Step 13, postvaccinal for step 12 supernatant liquor B3 entered shaking table 25-28 DEG C, 150-200rpm concussion cultivates 48-72 hour; Standing 30-60min after cultivation completes, discards 30ml supernatant liquor, gets precipitation 5ml and is inoculated into continuation cultivation in supernatant liquor B2;
Step 14, postvaccinal for step 13 supernatant liquor B2 entered shaking table 25-28 DEG C, 150-200rpm concussion cultivates 48-72 hour; Standing 30-60min after cultivation completes, discards 30ml supernatant liquor, gets precipitation 5ml and is inoculated into continuation cultivation in supernatant liquor B1;
Step 15, postvaccinal for step 14 supernatant liquor B1 entered shaking table 25-28 DEG C, 150-200rpm concussion cultivates 48-72 hour; Standing 30-60min after cultivation completes, discards 30ml supernatant liquor, when sample is tamed after cultivation through step supernatant liquor B1, get precipitation 1ml, be inoculated in the 3rd enrichment pipe, the pipe lid of the 3rd enrichment pipe is tightened, keep flat into shaking table 28-35 DEG C, 150-200rpm concussion cultivation 24-48 hour; By the 3rd enrichment pipe refrigeration or freezen protective after cultivation, after this domestication, bacterial classification is aerobic domesticated strain.
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| Title |
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| 于霞等: "煤化工废水COD高效降解菌降解性能研究", 《浙江农业科学》 * |
| 李扬等: "煤化工废水处理技术研究进展", 《山西水利科技》 * |
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