CN102559542A - Strain Pandoraea sp. for removing ammonia nitrogen in sewage under low-temperature condition and isolated culture method - Google Patents

Strain Pandoraea sp. for removing ammonia nitrogen in sewage under low-temperature condition and isolated culture method Download PDF

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CN102559542A
CN102559542A CN2011104162891A CN201110416289A CN102559542A CN 102559542 A CN102559542 A CN 102559542A CN 2011104162891 A CN2011104162891 A CN 2011104162891A CN 201110416289 A CN201110416289 A CN 201110416289A CN 102559542 A CN102559542 A CN 102559542A
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ammonia nitrogen
sewage
pandoraea
bacterium
nitrite bacteria
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CN102559542B (en
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胡春萍
杨蛟云
刘智晓
熊欢欢
张伟
沈旺
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Shouchuang Aihua (tianjin) Municipal & Environmental Engineering Co Ltd
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Shouchuang Aihua (tianjin) Municipal & Environmental Engineering Co Ltd
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    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02WCLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
    • Y02W10/00Technologies for wastewater treatment
    • Y02W10/10Biological treatment of water, waste water, or sewage

Abstract

The invention discloses a strain Pandoraea sp. for removing ammonia nitrogen in sewage under a low-temperature condition and an isolated culture method. The strain Pandoraea sp.-2CGMCC No. 5284 for removing the ammonia nitrogen in the sewage under the low-temperature condition has the capacity of removing the ammonia nitrogen in the sewage under the low-temperature aerobic condition. Experimental results show that: although the strain is obviously inhibited by a low-temperature environment, the strain Pandoraea sp.-2CGMCC No. 5284 for removing the ammonia nitrogen in the sewage under the low-temperature condition has an obvious reinforcing effect of removing the ammonia nitrogen at the low temperature of 10 DEG C, the removal rate of the ammonia nitrogen is improved by 10 percent, and the highest concentration of the effluent ammonia nitrogen is 5.1mg/L and superior to the effluent standard of the first grade A (when the water temperature is less than or equal to 12 DEG C, the control index of the first grade A standard is 8mg/L).

Description

Coldcondition goes down except that the bacterium Pandoraea sp. and the isolation cultivation method of ammonia nitrogen in the sewage
Technical field
The invention belongs to sewage treatment area, relate to a strain coldcondition and go down except that the bacterium Pandoraea sp. and the isolation cultivation method of ammonia nitrogen in the sewage.
Background technology
The reason that low temperature produces great effect to the biological treatment ability is because the reduction of temperature has influenced microbial growth and metabolism and made that change has taken place microbial population in the Sewage treatment systems.In temperature decline process; The metabolic activity of mesophilic bacteria descends gradually; Just lose activity during to certain temperature; And the quantity of psychrophile that is suitable for low temperature environment be because self physiological property and the restraining effect of various ecological factors quantitatively can't reach the status of dominant microflora, thereby causes sewage work's effluent quality in winter to descend.Therefore need badly a kind of can be in the low-temperature aerobic condition be gone down except that sewage the bacterial strain of ammonia nitrogen.
Summary of the invention
The objective of the invention is to overcome the deficiency of prior art, provide a kind of coldcondition to go down except that the bacterium Pandoraea sp. of ammonia nitrogen in the sewage
Second purpose of the present invention provides a kind of coldcondition and goes down except that the isolation cultivation method of the bacterium Pandoraea sp. of ammonia nitrogen in the sewage.
The 3rd purpose of the present invention provides a kind of coldcondition and goes down except that the purposes of the bacterium Pandoraea sp. of ammonia nitrogen in the sewage.
Coldcondition goes down except that the inferior Huang of the bacterium Pandoraea sp. of ammonia nitrogen in the sewage-2CGMCC No.5284, and it has the ability of ammonia nitrogen in the low-temperature aerobic condition is gone down except that sewage.
Coldcondition goes down except that the purposes of the inferior Huang of the bacterium Pandoraea sp. of ammonia nitrogen in the sewage-2CGMCC No.5284 ammonia nitrogen in the low-temperature aerobic condition is gone down except that sewage.
Said low temperature is preferably 8-12 ℃.
Coldcondition goes down except that the isolation cultivation method of the bacterium Pandoraea sp. of ammonia nitrogen in the sewage, it is characterized in that comprising the steps:
(1) first kind of Nitrite bacteria nutrient solution put into lab scale sbr reactor device, sludge of sewage treatment plant is inoculated in said first kind of Nitrite bacteria nutrient solution, at 8~12 ℃, cultured continuously 4-6 days; Take out supernatant, add second kind of Nitrite bacteria nutrient solution, at 8~12 ℃; Cultured continuously 4-6 days, take out supernatant, add the third Nitrite bacteria nutrient solution; At 8~12 ℃, cultured continuously 4-6 days, enrichment, domestication psychrotropic bacteria;
(2) get the mud that 10mL step (1) obtains, add in the 250ml Erlenmeyer flask that 90mL sterile purified water and 20~30 sterilization granulated glass spherees have been housed, vibration 10~30min makes bacterial cell fully be discharged in the supernatant; Leave standstill 20~40min, get the adding of 1mL supernatant and be equipped with in the test tube of 9mL sterile distilled water, obtain 10 -2Bacteria suspension, take turns doing serial dilution as stated above, accomplish 10 always -8Till;
(3) choosing gradient is 10 -5~10 -8Bacteria suspension, get 1mL respectively and be inoculated in the Nitrite bacteria solid medium, each extent of dilution is done 3 parallel appearance, numbering; Under the condition of aerobic, preserve moisture for 10 ℃ and cultivated for 2~3 weeks;
(4) choosing colony is big in the Nitrite bacteria solid medium flat board from step (3), single bacterium colony that form is different, and line separates once more in Nitrite bacteria solid medium flat board; Repeating plate streaking separates 3-4 time; Make the homomorphosis of bacterium colony in microscope on the flat board, single, big or small similar single bacterium colony that is; Called after is inferior yellow-2, through be accredited as coldcondition go down except that sewage in the bacterium Pandoraea sp. of ammonia nitrogen.
Said first kind of Nitrite bacteria nutrient solution is made up of following component: NH 4Cl 0.0764g, sodium acetate 2g, MgSO 47H 2O0.05g, K 2HPO 40.2g, NaCl 0.12g, MnSO 4H 2O 0.01g, FeSO 40.01g adding distil water is to 1000mL.
Said second kind of Nitrite bacteria nutrient solution is made up of following component: NH 4Cl 0.191g, sodium acetate 2g, MgSO 47H 2O0.05g, K 2HPO 40.2g, NaCl 0.12g, MnSO 4H 2O 0.01g, FeSO 40.01g adding distil water is to 1000mL.
Said the third Nitrite bacteria nutrient solution is made up of following component: NH 4Cl 0.382g, sodium acetate 2g, MgSO 47H 2O0.05g, K 2HPO 40.2g, NaCl 0.12g, MnSO 4H 2O 0.01g, FeSO 40.01g adding distil water is to 1000mL.
Said Nitrite bacteria solid medium is made up of following component: NH 4Cl 0.0764g, sodium acetate 2g, MgSO 47H 2O0.05g, K 2HPO 40.2g, NaCl 0.12g, MnSO 4H 2O 0.01g, FeSO 40.01g, agar 20g, adding distil water is to 1000mL.
Experimental result shows: although receive the obvious inhibition of low temperature environment; Add a kind of coldcondition of the present invention go down except that the inferior Huang of the bacterium Pandoraea sp. of ammonia nitrogen in the sewage-2CGMCC No.5284 under 10 ℃ of low temperature; Ammonia nitrogen removal is had tangible strengthening effect, and ammonia nitrogen removal frank has improved 10%, and the water outlet ammonia nitrogen concentration is up to 5.1mg/L; What be superior to one-level A goes out water quality standard (water temperature≤12 ℃ time, the controlling index of one-level A standard is 8mg/L).
Description of drawings
Fig. 1 goes down except that the domestication separation and Culture schema of the bacterium Pandoraea sp. of ammonia nitrogen in the sewage for a kind of coldcondition.
Fig. 2 is inferior yellow-2 ammonia nitrogen removals experiment.
Fig. 3 is Pandoraea sp. (inferior yellow-a 2) method for determining bacteria flow process.
Embodiment
Through specific embodiment the present invention is further described below.
Embodiment 1
Coldcondition goes down except that the isolation cultivation method of the bacterium Pandoraea sp. of ammonia nitrogen in the sewage, comprises the steps:
(1) first kind of Nitrite bacteria nutrient solution put into lab scale sbr reactor device, recorded in the said first kind of Nitrite bacteria nutrient solution of sludge seeding of village's WWT plant aeration tank in Tianjin, at 10 ℃, cultured continuously 5 days; Take out supernatant, add second kind of Nitrite bacteria nutrient solution, at 10 ℃; Cultured continuously 5 days is taken out supernatant, adds the third Nitrite bacteria nutrient solution; At 10 ℃, cultured continuously 5 days, enrichment, domestication psychrotropic bacteria;
(2) get the mud that 10mL step (1) obtains, add in the 250ml Erlenmeyer flask that 90mL sterile purified water and 30 sterilization granulated glass spherees have been housed, vibration 20min makes bacterial cell fully be discharged in the supernatant; Leave standstill 30min, get the adding of 1mL supernatant and be equipped with in the test tube of 9mL sterile distilled water, obtain 10 -2Bacteria suspension, take turns doing serial dilution as stated above, accomplish 10 always -8Till;
(3) choosing gradient is 10 -5~10 -8Bacteria suspension, get 1mL respectively and be inoculated in the Nitrite bacteria solid medium, each extent of dilution is done 3 parallel appearance, numbering; Under the condition of aerobic, preserve moisture for 10 ℃ and cultivated for 3 weeks;
(4) choosing colony is big in the Nitrite bacteria solid medium flat board from step (3), splashes into the different single bacterium colony of form that griess reagent reddens, and line separates once more in Nitrite bacteria solid medium flat board; Repeating plate streaking separates 3 times; Make the homomorphosis of bacterium colony in microscope on the flat board, single, big or small similar single bacterium colony that is; Called after is inferior yellow-2, through be accredited as coldcondition go down except that sewage in the bacterium Pandoraea sp. of ammonia nitrogen.
Embodiment 2
First kind of Nitrite bacteria nutrient solution is made up of following component: NH 4Cl 0.0764g, sodium acetate 2g, MgSO 47H 2O0.05g, K 2HPO 40.2g, NaCl 0.12g, MnSO 4H 2O 0.01g, FeSO 40.01g adding distil water is to 1000mL.
Embodiment 3
Second kind of Nitrite bacteria nutrient solution is made up of following component: NH 4Cl 0.191g, sodium acetate 2g, MgSO 47H 2O 0.05g, K 2HPO 40.2g, NaCl 0.12g, MnSO 4H 2O 0.01g, FeSO 40.01g adding distil water is to 1000mL.
Embodiment 4
The third Nitrite bacteria nutrient solution is made up of following component: NH 4Cl 0.382g, sodium acetate 2g, MgSO 47H 2O 0.05g, K 2HPO 40.2g, NaCl 0.12g, MnSO 4H 2O 0.01g, FeSO 40.01g adding distil water is to 1000mL.
Embodiment 5
The Nitrite bacteria solid medium is made up of following component: NH 4Cl 0.0764g, sodium acetate 2g, MgSO 47H 2O 0.05g, K 2HPO 40.2g, NaCl 0.12g, MnSO 4H 2O 0.01g, FeSO 40.01g, agar 20g, adding distil water is to 1000mL.
Embodiment 6
The preparation of griess reagent
1, Sulphanilic Acid reagent (A liquid): 0.5 gram Sulphanilic Acid is dissolved in 150 milliliter 20% the dilute acetic acid solution, is stored in brown bottle, refrigerates subsequent use.
2, alpha-naphthylamine reagent (B liquid): 0.5 gram alpha-naphthylamine is added in the dilute acetic acid solution of 20 ml distilled waters and 150 milliliter 20%, is stored in brown bottle, refrigerates subsequent use.
3, the use liquid of griess reagent: the A liquid of getting equal proportion mixes and can use with B liquid.
Embodiment 7
The bacterium Pandoraea sp. that coldcondition of the present invention goes down except that ammonia nitrogen in the sewage has following Microbiological Characteristics:
1. morphological characteristic
Examine under a microscope, for shaft-like, long 1.4 μ m, wide 0.28 μ m; On the Nitrite bacteria solid medium, form the oyster white bacterium colony.
2. cultivate and learn characteristic
Under 10 ℃, when in the Nitrite bacteria solid medium, cultivating, Pandoraea sp. bacterial strain of the present invention has following characteristic.
Smooth surface is moistening, the neat in edge rule, and individual small rounded, thalline is creamy white.
3. physiological property
Aerobic condition goes down except that the gram negative bacterium of ammonia nitrogen in the sewage, catalase is positive, methyl red test is negative, not hydrolyzed starch, liquefy gelatin, edwardsiella hoshinae, growth pH value are not 6~8,5~30 ℃ of growth temperatures.
Gram Catalase Methyl red The starch hydrolysis The gelatin test Indole test p?H Temperature
G - + - - ?- - 6~8 5~30
The 16s Sequence Identification of bacterium of the present invention
16SrDNA is the sequence that bacterium class prokaryotic organism are very guarded in very long evolutionary process; Playing the part of important role in the taxonomy of mikrobe in modern times; As the most frequently used phyletic evolution tagged molecule of bacterial flora structural analysis, it is more efficient and convenient to carry out structural analysis of microbial community with 16S rDNA.
Carrying out Bacteria Identification with the 16s sequence is undertaken by Fig. 3.
Beef-protein medium among Fig. 3: Carnis Bovis seu Bubali cream 3g, peptone 10g, sodium-chlor 5g, adding distil water are to 1000mL, and pH 7.0~7.2
The LB substratum: tryptone 10g, yeast extract 5g, NaCl 10g, adding distil water is to 1000ml.
The 16s sequence that the present invention records is as follows:
ACGCTGGCGGCATGCCTTACACATGCAAGTCGAACGGCAGCACGGGTGCTTGCACCTGGTGGCGAGT
GGCGAACGGGTGAGTAATACATCGGAATGTACCTTGTAGTGGGGGATAGCTCGGCGAAAGCCGGATT
AATACCGCATACGCTCTGAGGAGGAAAGCGGGGGACCTTCGGGCCTCGCGCTACAAGAGCAGCCGAT
GTCAGATTAGCTAGTTGGTGGGGTAAAAGCTCACCAAGGCGACGATCTGTAGCTGGTCTGAGAGGAC
GACCAGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATTTTGG
ACAATGGGCGAAAGCCTGATCCAGCAATGCCGCGTGTGTGAAGAAGGCCTTCGGGTTGTAAAGCACT
TTTGTCCGGAAAGAAATCCTCTGGGTTAATACCTCGGGGGGATGACGGTACCGGAAGAATAAGCACC
GGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTA
AAGCGTGCGCAGGCGGTTTTGTAAGACGGATGTGAAATCCCCGGGCTTAACCTGGGAACTGCATTCG
TGACTGCAAGGCTAGAGTATGGCAGAGGGGGGTAGAATTCCACGTGTAGCAGTGAAATGCGTAGAGA
TGTGGAGGAATACCGATGGCGAAGGCAGCCCCCTGGGCCAATACTGACGCTCATGCACGAAAGCGTG
GGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCCTAAACGATGTCAACTAGTTGTTGGGGATT
CATTTCCTTAGTAACGAAGCTAACGCGTGAAGTTGACCGCCTGGGGAGTACGGTCGCAAGATTAAAA
CTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGATGATGTGGATTAATTCGATGCAACGCGAAA
AACCTTACCTACCCTTGACATGTACGGAATCCTGCTGAGAGGTGGGAGTGCTCGAAAGAGAACCGTA
ACACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCG
CAACCCTTGTCCTTAGTTGCTACGCAAGAGCACTCTAGGGAGACTGCCGGTGACAAACCGGAGGAAG
GTGGGGATGACGTCAAGTCCTCATGGCCCTTATGGGTAGGGCTTCACACGTCATACAATGGTCGGTAC
AGAGGGCTGCCAAACCGCGAGGTGGAGCTAACCCCAGAAAACCGATCGTAGTCCGGATCGCAGTCT
GCAACTCGACTGCGTGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGTCGCGGTGAATACGTTCC
CGGGTCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTTTGCCAGAAGTAGGTAGCCTAACCGT
AAGGAGGGCGCTTACCACGGCAGGATTCATGACTGGG
Learn characteristic according to mentioned microorganism; Called after inferior yellow-2; Classification called after bacterium (Pandoraea sp.) is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on September 22nd, 2011, and deposit number is CGMCCNo.5284, and the preservation address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City; Institute of Microorganism, Academia Sinica, and survived.
Embodiment 8
The degradation effect of ammonia nitrogen in inferior yellow-2 pairs of sewage
In the beaker of 1L, add the 4th kind of Nitrite bacteria nutrient solution of 500m L (initial ammonia nitrogen concentration is 30mg/L), add an amount of inferior yellowly-2, maintenance pH is 7.5, its degraded situation of measuring 72 hours to ammonia nitrogen, the result is as shown in Figure 2.
Can be found out that by Fig. 2 through 72 hours degraded, ammonia nitrogen concentration was 1.2mg/L in the solution, ammonia nitrogen is removed basically, and clearance is 96%, proves that inferior yellow-2 pairs of ammonia nitrogens among the present invention have good removal ability.
The 4th kind of Nitrite bacteria nutrient solution formed: first kind of Nitrite bacteria nutrient solution is made up of following component: NH 4Cl0.1146g, sodium acetate 2g, MgSO 47H 2O 0.05g, K 2HPO 40.2g, NaCl 0.12g, MnSO 4H 2O 0.01g, FeSO 40.01g adding distil water is to 1000mL.
Embodiment 9
Coldcondition goes down except that the isolation cultivation method of the bacterium Pandoraea sp. of ammonia nitrogen in the sewage, comprises the steps:
(1) first kind of Nitrite bacteria nutrient solution put into lab scale sbr reactor device, sludge of sewage treatment plant is inoculated in said first kind of Nitrite bacteria nutrient solution, at 8 ℃, cultured continuously 6 days; Take out supernatant, add second kind of Nitrite bacteria nutrient solution, at 8 ℃; Cultured continuously 6 days is taken out supernatant, adds the third Nitrite bacteria nutrient solution; At 8 ℃, cultured continuously 6 days, enrichment, domestication psychrotropic bacteria;
(2) get the mud that 10mL step (1) obtains, add in the 250ml Erlenmeyer flask that 90mL sterile purified water and 20 sterilization granulated glass spherees have been housed, vibration 10~30min makes bacterial cell fully be discharged in the supernatant; Leave standstill 20~40min, get the adding of 1mL supernatant and be equipped with in the test tube of 9mL sterile distilled water, obtain 10 -2Bacteria suspension, take turns doing serial dilution as stated above, accomplish 10 always -8Till;
(3) choosing gradient is 10 -5~10 -8Bacteria suspension, get 1mL respectively and be inoculated in the Nitrite bacteria solid medium, each extent of dilution is done 3 parallel appearance, numbering; Under the condition of aerobic, preserve moisture for 10 ℃ and cultivated for 3 weeks;
(4) choosing colony is big in the Nitrite bacteria solid medium flat board from step (3); Single bacterium colony that form is different, line separates once more in Nitrite bacteria solid medium flat board, repeats plate streaking and separates 3 times; Make the homomorphosis of bacterium colony in microscope on the flat board; Single, size is similar to be single bacterium colony, through be accredited as coldcondition go down except that sewage in the bacterium Pandoraea sp. of ammonia nitrogen.
Embodiment 10
Coldcondition goes down except that the isolation cultivation method of the bacterium Pandoraea sp. of ammonia nitrogen in the sewage, comprises the steps:
(1) first kind of Nitrite bacteria nutrient solution put into lab scale sbr reactor device, sludge of sewage treatment plant is inoculated in said first kind of Nitrite bacteria nutrient solution, at 12 ℃, cultured continuously 4 days; Take out supernatant, add second kind of Nitrite bacteria nutrient solution, at 12 ℃; Cultured continuously 4 days is taken out supernatant, adds the third Nitrite bacteria nutrient solution; At 12 ℃, cultured continuously 4 days, enrichment, domestication psychrotropic bacteria;
(2) get the mud that 10mL step (1) obtains, add in the 250ml Erlenmeyer flask that 90mL sterile purified water and 30 sterilization granulated glass spherees have been housed, vibration 10~30min makes bacterial cell fully be discharged in the supernatant; Leave standstill 20~40min, get the adding of 1mL supernatant and be equipped with in the test tube of 9mL sterile distilled water, obtain 10 -2Bacteria suspension, take turns doing serial dilution as stated above, accomplish 10 always -8Till;
(3) choosing gradient is 10 -5~10 -8Bacteria suspension, get 1mL respectively and be inoculated in the Nitrite bacteria solid medium, each extent of dilution is done 3 parallel appearance, numbering; Under the condition of aerobic, preserve moisture for 10 ℃ and cultivated for 2 weeks;
(4) choosing colony is big in the Nitrite bacteria solid medium flat board from step (3); Single bacterium colony that form is different, line separates once more in Nitrite bacteria solid medium flat board, repeats plate streaking and separates 4 times; Make the homomorphosis of bacterium colony in microscope on the flat board; Single, size is similar to be single bacterium colony, through be accredited as coldcondition go down except that sewage in the bacterium Pandoraea sp. of ammonia nitrogen.
The experiment proof; Embodiment 9 and embodiment 10 through be accredited as coldcondition go down except that sewage in the bacterium Pandoraea sp. of isolation cultivation method separation and Culture of bacterium Pandoraea sp. of ammonia nitrogen the effect of good removal ammonia nitrogen is also arranged, clearance is respectively 95.8% and 95.4%.

Claims (8)

1. coldcondition goes down except that the inferior Huang of the bacterium Pandoraea sp. of ammonia nitrogen in the sewage-2CGMCC No.5284, and it has the ability of ammonia nitrogen in the low-temperature aerobic condition is gone down except that sewage.
2. the coldcondition of claim 1 goes down except that the purposes of the inferior Huang of the bacterium Pandoraea sp. of ammonia nitrogen in the sewage-2CGMCC No.5284 ammonia nitrogen in the low-temperature aerobic condition is gone down except that sewage.
3. purposes according to claim 2 is characterized in that said low temperature is 8-12 ℃.
4. coldcondition goes down except that the isolation cultivation method of the bacterium Pandoraea sp. of ammonia nitrogen in the sewage, it is characterized in that comprising the steps:
(1) first kind of Nitrite bacteria nutrient solution put into lab scale sbr reactor device, sludge of sewage treatment plant is inoculated in said first kind of Nitrite bacteria nutrient solution, at 8~12 ℃, cultured continuously 4-6 days; Take out supernatant, add second kind of Nitrite bacteria nutrient solution, at 8~12 ℃; Cultured continuously 4-6 days, take out supernatant, add the third Nitrite bacteria nutrient solution; At 8~12 ℃, cultured continuously 4-6 days, enrichment, domestication psychrotropic bacteria;
(2) get the mud that 10mL step (1) obtains, add in the 250ml Erlenmeyer flask that 90mL sterile purified water and 20~30 sterilization granulated glass spherees have been housed, vibration 10~30min makes bacterial cell fully be discharged in the supernatant; Leave standstill 20~40min, get the adding of 1mL supernatant and be equipped with in the test tube of 9mL sterile distilled water, obtain 10 -2Bacteria suspension, take turns doing serial dilution as stated above, accomplish 10 always -8Till;
(3) choosing gradient is 10 -5~10 -8Bacteria suspension, get 1mL respectively and be inoculated in the Nitrite bacteria solid medium, each extent of dilution is done 3 parallel appearance, numbering; Under the condition of aerobic, preserve moisture for 10 ℃ and cultivated for 2~3 weeks;
(4) choosing colony is big in the Nitrite bacteria solid medium flat board from step (3), single bacterium colony that form is different, and line separates once more in Nitrite bacteria solid medium flat board; Repeating plate streaking separates 3-4 time; Make the homomorphosis of bacterium colony in microscope on the flat board, single, big or small similar single bacterium colony that is; Called after is inferior yellow-2, through be accredited as coldcondition go down except that sewage in the bacterium Pandoraea sp. of ammonia nitrogen.
5. coldcondition according to claim 4 goes down except that the isolation cultivation method of the bacterium Pandoraea sp. of ammonia nitrogen in the sewage, it is characterized in that said first kind of Nitrite bacteria nutrient solution be made up of following component: NH 4Cl 0.0764g, sodium acetate 2g, MgSO 47H 2O 0.05g, K 2HPO 40.2g, NaCl 0.12g, MnSO 4H 2O 0.01g, FeSO 40.01g adding distil water is to 1000mL.
6. coldcondition according to claim 4 goes down except that the isolation cultivation method of the bacterium Pandoraea sp. of ammonia nitrogen in the sewage, it is characterized in that said second kind of Nitrite bacteria nutrient solution be made up of following component: NH 4Cl 0.191g, sodium acetate 2g, MgSO 47H 2O 0.05g, K 2HPO 40.2g, NaCl 0.12g, MnSO 4H 2O 0.01g, FeSO 40.01g adding distil water is to 1000mL.
7. coldcondition according to claim 4 goes down except that the isolation cultivation method of the bacterium Pandoraea sp. of ammonia nitrogen in the sewage, it is characterized in that said the third Nitrite bacteria nutrient solution is made up of following component: NH 4Cl 0.382g, sodium acetate 2g, MgSO 47H 2O 0.05g, K 2HPO 40.2g, NaCl 0.12g, MnSO 4H 2O 0.01g, FeSO 40.01g adding distil water is to 1000mL.
8. coldcondition according to claim 4 goes down except that the isolation cultivation method of the bacterium Pandoraea sp. of ammonia nitrogen in the sewage, it is characterized in that said Nitrite bacteria solid medium is made up of following component: NH 4Cl 0.0764g, sodium acetate 2g, MgSO 47H 2O 0.05g, K 2HPO 40.2g, NaCl 0.12g, MnSO 4H 2O 0.01g, FeSO 40.01g, agar 20g, adding distil water is to 1000mL.
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CN103756928A (en) * 2013-11-26 2014-04-30 浙江大学 Bacterial strain for degradation of p-xylene and culture method and application thereof

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CN103319000A (en) * 2013-07-15 2013-09-25 北京大学 Enrichment of low temperature resistant autotrophic nitrification microbial agent and its application in wastewater treatment
CN103319000B (en) * 2013-07-15 2014-10-08 北京大学 Enrichment of low temperature resistant autotrophic nitrification microbial agent and its application in wastewater treatment
CN103756928A (en) * 2013-11-26 2014-04-30 浙江大学 Bacterial strain for degradation of p-xylene and culture method and application thereof

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