CN105368914A - Compound microorganism screening method for degrading domestic sewage - Google Patents

Compound microorganism screening method for degrading domestic sewage Download PDF

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Publication number
CN105368914A
CN105368914A CN201510927152.0A CN201510927152A CN105368914A CN 105368914 A CN105368914 A CN 105368914A CN 201510927152 A CN201510927152 A CN 201510927152A CN 105368914 A CN105368914 A CN 105368914A
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China
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yeast
cgmccno
sewage
treatment
sanitary sewage
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乔长晟
李坤荣
孙雨
钟海蛟
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Tianjin Peiyang Biotrans Biotech Co Ltd
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Tianjin Peiyang Biotrans Biotech Co Ltd
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Abstract

The invention discloses a compound microorganism screening method for degrading domestic sewage. The method comprises the following steps: inoculating to-be-screened compound microorganisms in urban sewage for biological treatment which lasts for 5; respectively analyzing total volatile organic compounds (TVOC) and odor ingredients in the domestic sewage before and after the biological treatment by virtue of an electronic nose; and determining final compound microorganism for the efficient degradation of the domestic sewage by taking the reduction and content reducing degree of the TVOC and odor materials as indexes for screening a dominant compound strain which is used for the efficient degradation of the domestic sewage. Through the method, change in the TVOC and odor ingredients in the sewage before and after the treatment on the compound strain can be precisely determined, so that obtained change information related to the TVOC and odor ingredients in the sewage is comprehensive; and the method, for compound strain screening, has the advantages of being sensitive and rapid, and being comprehensive in water quality information.

Description

The complex microorganism screening method of degraded sanitary sewage
Technical field
The present invention relates to the technical field of sanitary sewage disposal, is a kind of complex microorganism screening method of sanitary sewage of degrading specifically.
Background technology
China's shortage of water resources, water resources ownership per capita is lower than world average level, and urbanite water consumption amount strengthens gradually, for quality of water environment concern and require day by day improve.Therefore process is carried out to city domestic sewage very urgent.In city domestic sewage, BOD, COD, SS, oil and the content of nitrogen and phosphorous are all higher, can volatilize a large amount of stench class materials: NH3 and H2S in the process of process.
The proportion that in sanitary sewage, VOCs accounts for is larger, and it has stable chemical nature, boiling point is low, density is large, viscosity is little, be insoluble in the characteristic of water, some or suspect carcinogen matter.VOCs (volatile organic contaminant) is a kind of important pollutent, irritant, has and smells taste, can have a strong impact on the central nervous system of people and Digestive tract thus harm humans is healthy.Thus in sanitary sewage disposal process, the detection of TVOC and stench is seemed particularly important.
Electric nasus system can be simulated Biologic Olfaction system, and the gas signal in sewage is converted to electrical signal, carries out qualitative and quantitative analysis to VOCs, thus realizes for sanitary sewage water quality Real-Time Monitoring.Biologic treating technique utilizes the metabolism of microorganism that organic pollutant is decomposed into CO 2and water, or organic acid and alcohol etc., thus reach the object of purifying waste water.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of complex microorganism screening method of sanitary sewage of degrading.
The technical scheme that the present invention takes for the technical problem existed in solution known technology is:
The biological treatment carrying out by a definite date 5d in composite microbial inoculant to be screened to city domestic sewage of the present invention, Electronic Nose is utilized to analyze respectively the TVOC in the sanitary sewage before and after biological treatment and stench constituents, using TVOC and the reduction of stench class material and the minimizing degree of content as the index of screening efficient degradation sanitary sewage super hybrid bacteria strain, to determine the complex microorganism of final efficient degradation sanitary sewage.
The present invention can also adopt following technical measures:
Concrete steps based on the complex microorganism screening method of Electronic Nose are as follows:
(1) actication of culture: each bacterial classification activates respectively, lines respectively by it on corresponding solid medium;
(2) seed liquor preparation: the bacterium colony of picking activation, is inoculated in respectively in each self-corresponding liquid nutrient medium and cultivates;
(3) with Bioscreen full-automatic growth curve determinator, each bacterial classification is carried out to the mensuration of growth curve, determine that thalli growth arrives the time of logarithmic phase, as the cultivation period of seed liquor when preparing composite microbial bacteria liquid.
(4) will the seed liquor of logarithmic phase be cultured to according to being inoculated in sanitary sewage respectively, subsequently on shaking table with 20 DEG C, the condition of 110rpm carries out the purifying treatment of 5 days by a definite date.
(5) utilize Electronic Nose to analyze with the VOCs in the sewage before and after the process of composite microbial bacteria liquid and stench, by through complex microorganism before and after treatment after the sewage water bath with thermostatic control shaking table of putting into 60 DEG C respectively heat 1h and make the gas of head space and liquid reach balance; Gas is set to 150ml/min through the flow velocity of sensor, and inject time is 60s; Detected result data before and after treatment can be obtained, data are analyzed after dimension-reduction treatment and PCA are analyzed.
Choose the yeast of preservation, bacterium and the milk-acid bacteria bacterial classification that sets out as screening respectively, concrete steps are as follows:
(1) actication of culture: the multiple yeast preserved on 4 DEG C of inclined-planes, bacterium and milk-acid bacteria activate respectively, it are lined on corresponding solid medium respectively, and yeast cultivates 2-3d in 30 DEG C, and bacterium cultivates 1d in 37 DEG C, and milk-acid bacteria 37 DEG C cultivates 2-3d;
(2) seed liquor preparation: the bacterium colony of picking activation, is inoculated in respective liquid nutrient medium respectively, yeast in 30 DEG C, 180rpm cultivates 2d, bacterium and milk-acid bacteria in 37 DEG C, 200rpm cultivates 2d;
Seed culture medium consists of:
Microzyme culture medium is YEPD substratum, and in this substratum, each concentration of component is: peptone 2g/L, yeast extract paste 1g/L, glucose 2g/L, PH6.0,121 DEG C of sterilizing 20min;
Bacteria culture medium is broth culture, and in this substratum, each concentration of component is: peptone 10g/L, extractum carnis 5g/L, NaCl5g/L, PH7.2-7.4,121 DEG C of sterilizing 20min;
Lactic acid bacteria culturing medium is MRS substratum, and in this substratum, each concentration of component is: peptone 10g/L, beef powder 5g/L, yeast powder 4g/L, glucose 20g/L, K2HPO42g/L, anhydrous sodium acetate 5g/L, dibasic ammonium citrate 2g/L, magnesium sulfate 0.2g/L, manganous sulfate 0.05g/L, in addition, add POLYSORBATE 80 1.0ml, PH6.2-6.4,121 DEG C of sterilizing 15min;
(3) with Bioscreen full-automatic growth curve determinator, each yeast, bacterium and milk-acid bacteria are carried out to the mensuration of growth curve, determine that thalli growth arrives the time of logarithmic phase, as the cultivation period of seed liquor when preparing composite microbial bacteria liquid.
(4) will the seed liquor of logarithmic phase be cultured to according to being inoculated in sanitary sewage respectively, subsequently on shaking table with 20 DEG C, the condition of 110rpm carries out the purifying treatment of 5 days by a definite date.
(5) utilize Electronic Nose to analyze with the VOCs in the sewage before and after the process of composite microbial bacteria liquid and stench, by through complex microorganism before and after treatment after the sewage water bath with thermostatic control shaking table of putting into 60 DEG C respectively heat 1h and make the gas of head space and liquid reach balance; Gas is set to 150ml/min through the flow velocity of sensor, and inject time is 60s; Detected result data before and after treatment can be obtained, data are analyzed after dimension-reduction treatment and PCA are analyzed.
The composite microbial bacteria liquid be inoculated in sanitary sewage is following six kinds, and the different strain in each composite bacteria carries out proportioning in mass ratio, wherein yeast CGMCCNO.2.2076: yeast CGMCCNO.2.1554=1:1; Bacillus cereus CGMCCNO.1.234: Pseudomonas fluorescence=1:1; Yeast CGMCCNO.2.2076: Pseudomonas fluorescence: Lactobacterium acidophilum CGMCCNO.1.2912=1:1:1; Yeast CGMCCNO.2.2076: bacillus cereus CGMCCNO.1.234: Lactobacterium acidophilum=1:1:1; Yeast CGMCCNO.2.1554: Pseudomonas fluorescence: Lactobacterium acidophilum CGMCCNO.1.2912=1:1:1; Yeast CGMCCNO.2.1554: bacillus cereus CGMCCNO.1.234: Lactobacterium acidophilum CGMCCNO.1.2912=1:1:1, wherein Pseudomonas fluorescence is bought from Biological Engineering College of University Of Science and Technology Of Tianjin Microbiological Culture Collection storehouse, and all the other bacterial classifications are all purchased from China General Microbiological culture presevation administrative center.
The advantage that the present invention has and positively effect are:
In the complex microorganism screening method of degraded sanitary sewage of the present invention, the biological treatment of 5d by a definite date will be carried out in composite microbial inoculant to be screened to city domestic sewage, Electronic Nose is utilized to analyze respectively the TVOC in the sanitary sewage before and after biological treatment and stench constituents, using TVOC and the reduction of stench class material and the minimizing degree of content as the index of screening efficient degradation sanitary sewage super hybrid bacteria strain, to determine the complex microorganism of final efficient degradation sanitary sewage.Present method accurately can measure the changing conditions of TVOC and stench constituents in sewage before and after compound strain process, in the relevant sewage obtained, the change information of TVOC and stench constituents is comprehensive, for compound strain screening operation have sensitive, quick, the comprehensive advantage of water quality information.
Embodiment
Below by way of specific embodiment, the technical scheme in the present invention is described in detail:
The biological treatment carrying out by a definite date 5d in composite microbial inoculant to be screened to city domestic sewage of the present invention, Electronic Nose is utilized to analyze respectively the TVOC in the sanitary sewage before and after biological treatment and stench constituents, using TVOC and the reduction of stench class material and the minimizing degree of content as the index of screening efficient degradation sanitary sewage super hybrid bacteria strain, to determine the complex microorganism of final efficient degradation sanitary sewage.
Choose the yeast of preservation, bacterium and the milk-acid bacteria bacterial classification that sets out as screening respectively, concrete steps are as follows:
(1) actication of culture: the multiple yeast preserved on 4 DEG C of inclined-planes, bacterium and milk-acid bacteria activate respectively, it are lined on corresponding solid medium respectively, and yeast cultivates 2-3d in 30 DEG C, and bacterium cultivates 1d in 37 DEG C, and milk-acid bacteria 37 DEG C cultivates 2-3d;
(2) seed liquor preparation: the bacterium colony of picking activation, is inoculated in respective liquid nutrient medium respectively, yeast in 30 DEG C, 180rpm cultivates 2d, bacterium and milk-acid bacteria in 37 DEG C, 200rpm cultivates 2d;
Seed culture medium consists of:
Microzyme culture medium is YEPD substratum, and in this substratum, each concentration of component is: peptone 2g/L, yeast extract paste 1g/L, glucose 2g/L, PH6.0,121 DEG C of sterilizing 20min;
Bacteria culture medium is broth culture, and in this substratum, each concentration of component is: peptone 10g/L, extractum carnis 5g/L, NaCl5g/L, PH7.2-7.4,121 DEG C of sterilizing 20min;
Lactic acid bacteria culturing medium is MRS substratum, and in this substratum, each concentration of component is: peptone 10g/L, beef powder 5g/L, yeast powder 4g/L, glucose 20g/L, K2HPO42g/L, anhydrous sodium acetate 5g/L, dibasic ammonium citrate 2g/L, magnesium sulfate 0.2g/L, manganous sulfate 0.05g/L, in addition, add POLYSORBATE 80 1.0ml, PH6.2-6.4,121 DEG C of sterilizing 15min;
(3) with Bioscreen full-automatic growth curve determinator, each yeast, bacterium and milk-acid bacteria are carried out to the mensuration of growth curve, determine that thalli growth arrives the time of logarithmic phase, as the cultivation period of seed liquor when preparing composite microbial bacteria liquid.
(4) will the seed liquor of logarithmic phase be cultured to according to being inoculated in sanitary sewage respectively, subsequently on shaking table with 20 DEG C, the condition of 110rpm carries out the purifying treatment of 5 days by a definite date.
The composite microbial bacteria liquid be inoculated in sanitary sewage is following six kinds, and the different strain in each composite bacteria carries out proportioning in mass ratio, wherein yeast CGMCCNO.2.2076: yeast CGMCCNO.2.1554=1:1; Bacillus cereus CGMCCNO.1.234: Pseudomonas fluorescence=1:1; Yeast CGMCCNO.2.2076: Pseudomonas fluorescence: Lactobacterium acidophilum CGMCCNO.1.2912=1:1:1; Yeast CGMCCNO.2.2076: bacillus cereus CGMCCNO.1.234: Lactobacterium acidophilum=1:1:1; Yeast CGMCCNO.2.1554: Pseudomonas fluorescence: Lactobacterium acidophilum CGMCCNO.1.2912=1:1:1; Yeast CGMCCNO.2.1554: bacillus cereus CGMCCNO.1.234: Lactobacterium acidophilum CGMCCNO.1.2912=1:1:1, wherein Pseudomonas fluorescence is bought from Biological Engineering College of University Of Science and Technology Of Tianjin Microbiological Culture Collection storehouse, and all the other bacterial classifications are all purchased from China General Microbiological culture presevation administrative center.
(5) utilize Electronic Nose to analyze with the VOCs in the sewage before and after the process of composite microbial bacteria liquid and stench, by through complex microorganism before and after treatment after the sewage water bath with thermostatic control shaking table of putting into 60 DEG C respectively heat 1h and make the gas of head space and liquid reach balance; Gas is set to 150ml/min through the flow velocity of sensor, and inject time is 60s; Detected result data before and after treatment can be obtained, data are analyzed after dimension-reduction treatment and PCA are analyzed.
TVOC and the display of odor pollution analytical results: the TVOC contained in the sewage after two Yeasts liquid process is oxirane, alkene, ester class, siloxanes, and stench constituents is: amine; In sewage after two strain bacterial treatment, TVOC composition is siloxanes, oxirane, methyl acids, and stench constituents is amine, amino, chloride class; In sewage after yeast (CGMCCNO.2.1554), Pseudomonas fluorescence, the process of Lactobacterium acidophilum compound seed liquid, TVOC composition is alkene, alkane, siloxanes, lipid, and stench constituents is amine, amino, chlorine class; In sewage after yeast (CGMCCNO.2.1554), bacillus cereus, the process of Lactobacterium acidophilum composite bacteria liquid, TVOC composition is: siloxanes, oxirane, alkane, and stench constituents is chlorine class; In sewage after yeast (CGMCCNO.2.1554), Pseudomonas fluorescence, the process of Lactobacterium acidophilum composite bacteria liquid, TVOC composition is alkene, alkane, siloxanes, and stench constituents is amine; In sewage after yeast (CGMCCNO.2.1554), bacillus cereus, the process of Lactobacterium acidophilum composite bacteria liquid, TVOC composition is alcohols, alkene, ethers, and stench constituents is ammonia, containing amino.
The treatment process of carrying out when TVOC analyzes with stench constituents through composite bacteria liquid sewage before treatment Electronic Nose is identical with the treatment condition of sewage after composite bacteria liquid process, in untreated sewage, TVOC composition is oxirane, alkene, ester class, siloxanes, methyl acids, ethers, alcohols, lipid, siloxanes, and stench constituents is amine, containing amino, chlorine class.
Yeast (CGMCCNO.2.2076), bacillus cereus, the clearance of Lactobacterium acidophilum composite bacteria liquid to TVOC in sewage and stench constituents reach 90.24% and 76.31% respectively; Yeast (CGMCCNO.2.1554), Pseudomonas fluorescence, the Lactobacterium acidophilum composite bacteria liquid clearance to TVOC in sewage and stench constituents reaches 91.27% and 75.36% respectively.
Experiment shows that the removal effect of these two kinds of composite bacteria liquids to TVOC in sewage and stench constituents is better, Electronic Nose is utilized to analyze the TVOC in sewage before and after process and stench class material, thus filter out the good composite microbial bacteria liquid of effect of disposing of sewage, be respectively: yeast (CGMCCNO.2.2076): bacillus cereus (CGMCCNO.1.234): Lactobacterium acidophilum=1:1:1; Yeast (CGMCCNO.2.1554): Pseudomonas fluorescence: Lactobacterium acidophilum (CGMCCNO.1.2912)=1:1:1.
The above, it is only preferred embodiment of the present invention, not any pro forma restriction is done to the present invention, although the present invention with preferred embodiment openly as above, but, and be not used to limit the present invention, any those skilled in the art, do not departing within the scope of technical solution of the present invention, certainly the technology contents of announcement can be utilized to make a little change or modification, become the Equivalent embodiments of equivalent variations, in every case be the content not departing from technical solution of the present invention, according to any simple modification that technical spirit of the present invention is done above embodiment, equivalent variations and modification, all belong in the scope of technical solution of the present invention.

Claims (4)

1. the complex microorganism screening method of a sanitary sewage of degrading, it is characterized in that: the biological treatment of 5d by a definite date will be carried out in composite microbial inoculant to be screened to city domestic sewage, Electronic Nose is utilized to analyze respectively the TVOC in the sanitary sewage before and after biological treatment and stench constituents, using TVOC and the reduction of stench class material and the minimizing degree of content as the index of screening efficient degradation sanitary sewage super hybrid bacteria strain, to determine the complex microorganism of final efficient degradation sanitary sewage.
2. the complex microorganism screening method of degraded sanitary sewage according to claim 1, is characterized in that: the concrete steps based on the complex microorganism screening method of Electronic Nose are as follows:
(1) actication of culture: each bacterial classification activates respectively, lines respectively by it on corresponding solid medium;
(2) seed liquor preparation: the bacterium colony of picking activation, is inoculated in respectively in each self-corresponding liquid nutrient medium and cultivates;
(3) Bioscreen is used
Full-automatic growth curve determinator carries out the mensuration of growth curve to each bacterial classification, determine that thalli growth arrives the time of logarithmic phase, as the cultivation period of seed liquor when preparing composite microbial bacteria liquid.
(4) will the seed liquor of logarithmic phase be cultured to according to being inoculated in sanitary sewage respectively, subsequently on shaking table with 20 DEG C, the condition of 110rpm carries out the purifying treatment of 5 days by a definite date.
(5) utilize Electronic Nose to analyze with the VOCs in the sewage before and after the process of composite microbial bacteria liquid and stench, by through complex microorganism before and after treatment after the sewage water bath with thermostatic control shaking table of putting into 60 DEG C respectively heat 1h and make the gas of head space and liquid reach balance; Gas is set to 150ml/min through the flow velocity of sensor, and inject time is 60s; Detected result data before and after treatment can be obtained, data are analyzed after dimension-reduction treatment and PCA are analyzed.
3. the complex microorganism screening method of degraded sanitary sewage according to claim 2, is characterized in that: choose the yeast of preservation, bacterium and the milk-acid bacteria bacterial classification that sets out as screening respectively, concrete steps are as follows:
(1) actication of culture: the multiple yeast preserved on 4 DEG C of inclined-planes, bacterium and milk-acid bacteria activate respectively, it are lined on corresponding solid medium respectively, and yeast cultivates 2-3d in 30 DEG C, and bacterium cultivates 1d in 37 DEG C, and milk-acid bacteria 37 DEG C cultivates 2-3d;
(2) seed liquor preparation: the bacterium colony of picking activation, is inoculated in respective liquid nutrient medium respectively, yeast in 30 DEG C, 180rpm cultivates 2d, bacterium and milk-acid bacteria in 37 DEG C, 200rpm cultivates 2d;
Seed culture medium consists of:
Microzyme culture medium is YEPD substratum, and in this substratum, each concentration of component is: peptone 2g/L, yeast extract paste 1g/L, glucose 2g/L, PH6.0,121 DEG C of sterilizing 20min;
Bacteria culture medium is broth culture, and in this substratum, each concentration of component is: peptone 10g/L, extractum carnis 5g/L, NaCl5g/L, PH7.2-7.4,121 DEG C of sterilizing 20min;
Lactic acid bacteria culturing medium is MRS substratum, and in this substratum, each concentration of component is: peptone 10g/L, beef powder 5g/L, yeast powder 4g/L, glucose 20g/L, K2HPO42g/L, anhydrous sodium acetate 5g/L, dibasic ammonium citrate 2g/L, magnesium sulfate 0.2g/L, manganous sulfate 0.05g/L, in addition, add POLYSORBATE 80 1.0ml, PH6.2-6.4,121 DEG C of sterilizing 15min;
(3) Bioscreen is used
Full-automatic growth curve determinator carries out the mensuration of growth curve to each yeast, bacterium and milk-acid bacteria, determine that thalli growth arrives the time of logarithmic phase, as the cultivation period of seed liquor when preparing composite microbial bacteria liquid.
(4) will the seed liquor of logarithmic phase be cultured to according to being inoculated in sanitary sewage respectively, subsequently on shaking table with 20 DEG C, the condition of 110rpm carries out the purifying treatment of 5 days by a definite date.
(5) utilize Electronic Nose to analyze with the VOCs in the sewage before and after the process of composite microbial bacteria liquid and stench, by through complex microorganism before and after treatment after the sewage water bath with thermostatic control shaking table of putting into 60 DEG C respectively heat 1h and make the gas of head space and liquid reach balance; Gas is set to 150ml/min through the flow velocity of sensor, and inject time is 60s; Detected result data before and after treatment can be obtained, data are analyzed after dimension-reduction treatment and PCA are analyzed.
4. the complex microorganism screening method of degraded sanitary sewage according to claim 3, it is characterized in that: the composite microbial bacteria liquid be inoculated in sanitary sewage is following six kinds, different strain in each composite bacteria carries out proportioning in mass ratio, wherein yeast CGMCCNO.2.2076: yeast CGMCCNO.2.1554=1:1; Bacillus cereus CGMCCNO.1.234: Pseudomonas fluorescence=1:1; Yeast CGMCCNO.2.2076: Pseudomonas fluorescence: Lactobacterium acidophilum CGMCCNO.1.2912=1:1:1; Yeast CGMCCNO.2.2076: bacillus cereus CGMCCNO.1.234: Lactobacterium acidophilum=1:1:1; Yeast CGMCCNO.2.1554: Pseudomonas fluorescence: Lactobacterium acidophilum CGMCCNO.1.2912=1:1:1; Yeast CGMCCNO.2.1554: bacillus cereus CGMCCNO.1.234: Lactobacterium acidophilum CGMCCNO.1.2912=1:1:1, wherein Pseudomonas fluorescence is bought from Biological Engineering College of University Of Science and Technology Of Tianjin Microbiological Culture Collection storehouse, and all the other bacterial classifications are all purchased from China General Microbiological culture presevation administrative center.
CN201510927152.0A 2015-12-11 2015-12-11 Compound microorganism screening method for degrading domestic sewage Pending CN105368914A (en)

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Application publication date: 20160302