CN102424805B - Rhodococcus sp. for removing nitrite nitrogen from sewage at low temperature condition, and separation and culture method thereof - Google Patents
Rhodococcus sp. for removing nitrite nitrogen from sewage at low temperature condition, and separation and culture method thereof Download PDFInfo
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- CN102424805B CN102424805B CN2011104168934A CN201110416893A CN102424805B CN 102424805 B CN102424805 B CN 102424805B CN 2011104168934 A CN2011104168934 A CN 2011104168934A CN 201110416893 A CN201110416893 A CN 201110416893A CN 102424805 B CN102424805 B CN 102424805B
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- Y—GENERAL 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
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W10/00—Technologies for wastewater treatment
- Y02W10/10—Biological treatment of water, waste water, or sewage
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Abstract
The present invention discloses Rhodococcus sp. for removing nitrite nitrogen from sewage at a low temperature condition, and a separation and culture method thereof. The Rhodococcus sp. xiaoshen-1 CGMCC No.5277 for removing the nitrite nitrogen from the sewage at the low temperature condition has the function of removal of the nitrite nitrogen from the sewage at the low temperature and aerobic condition. The experimental results show that: the significant reinforcement effect of removal of the nitrite nitrogen in the sewage at the low temperature and especially at the temperature of 8-12 DEG C is provided despite the significant inhibition of the low temperature environment after the Rhodococcus sp. xiaoshen-1 CGMCC No.5277 of the present invention is added to the sewage, wherein the nitrite nitrogen removal rate in the sewage inoculated with the Rhodococcus sp. of the present invention is increased by about 10% compared to the nitrite nitrogen removal rate in the sewage without inoculation with the Rhodococcus sp. of the present invention.
Description
Technical field
The present invention relates to a strain rhodococcus and purposes and isolation cultivation method.
Background technology
The reason that low temperature produces great effect to the biological treatment ability is because the reduction of temperature has affected microbial growth and metabolism and made that change has occured microbial population in the Sewage treatment systems.In the drop in temperature process, the metabolic activity of mesophilic bacteria descends gradually, just lose activity during to certain temperature, and be suitable for low temperature environment psychrophile quantity since self physiological property and the restraining effect of various ecological factors, quantitatively can't reach the status of dominant microflora, thereby cause sewage work's effluent quality in winter to descend.Therefore need badly a kind ofly can go down except the bacterial strain of sewage Nitrite Nitrogen in the low-temperature aerobic condition.
Summary of the invention
The objective of the invention is to overcome the deficiencies in the prior art, a kind of rhodococcus of removing the Nitrite In Polluted Water nitrate nitrogen is provided.
Second purpose of the present invention provides a kind of purposes of removing the rhodococcus of Nitrite In Polluted Water nitrate nitrogen.
The 3rd purpose of the present invention provides a kind of isolation cultivation method of rhodococcus of removing nitrite nitrogen in sewage under low-temperature condition.
Technical scheme of the present invention is summarized as follows:
A kind of rhodococcus of removing nitrite nitrogen in sewage under low-temperature condition (Rhodococcus sp.) nitre is dark-1CGMCCNo.5277, and it has in the low-temperature aerobic condition to go down except the ability of sewage Nitrite Nitrogen.
A kind of rhodococcus of removing nitrite nitrogen in sewage under low-temperature condition (Rhodococcus sp.) nitre is dark-and 1CGMCCNo.5277 goes down except the purposes of sewage Nitrite Nitrogen in the low-temperature aerobic condition.Preferred 8～12 ℃ of described low temperature.
The isolation cultivation method of the rhodococcus of removing nitrite nitrogen in sewage under low-temperature condition comprises the steps:
(1) with the sludge seeding of 10～20mL sewage disposal plant aeration tank to the Erlenmeyer flask that 250mL Nitrate bacteria nutrient solution is housed, nitrogen concentration is 50mg/L in the Sodium Nitrite in the described Nitrate bacteria nutrient solution, at 8～12 ℃, shaking speed is to tame in 120～140rpm shaking table, when treating that the nitrite nitrogen degradation rate reaches 80～90%, leave standstill, remove supernatant liquor, with the sludge seeding of Erlenmeyer flask bottom to being equipped with in 250mL the second Nitrate bacteria nutrient solution, nitrite nitrogen concentration in described the second Nitrate bacteria nutrient solution is 150mg/L, at 8～12 ℃, in being the shaking table of 120～140rpm, rotating speed tames, when treating that the nitrite nitrogen degradation rate reaches 80～90%, namely obtain containing the Nitrate bacteria nutrient solution of low temperature resistant bacterium;
(2) get the nutrient solution that 10mL step (1) obtains, add oneself and be equipped with in the 250mL Erlenmeyer flask of 90mL sterile purified water and 20～30 sterilization granulated glass spherees, vibration 10～30min makes bacterial cell fully be discharged in the supernatant liquor; Leave standstill 20～40min, get the adding of 1mL supernatant liquor and be equipped with in the test tube of 9mL sterile distilled water, obtain 10
-2Bacteria suspension, take turns doing as stated above serial dilution, accomplish 10 always
-8Till;
(3) choosing gradient is 10
-5～10
-8Bacteria suspension, get respectively 1mL and be inoculated in the Nitrate bacteria solid medium, each extent of dilution is done 3 Duplicate Samples, the numbering; 10 ℃ of moisturizing 2～3 weeks of cultivation under the condition of aerobic;
(4) choosing colony is large in the Nitrate bacteria solid medium flat board from step (3), single bacterium colony that form is different, line separates again in Nitrate 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 nitre is dark-1, the rhodococcus through being accredited as removing nitrite nitrogen in sewage under low-temperature condition (Rhodococcus sp.).
Described Nitrate bacteria nutrient solution is comprised of following component: NaNO
20.25g, sodium acetate 2g, MgSO
47H
2O 0.05g, K
2HPO
40.2g, NaCl 0.12g, MnSO
44H
2O 0.01g, FeSO
40.01g, adding distil water is to 1000mL.
Described the second Nitrate bacteria nutrient solution is comprised of following component: NaNO
20.74g, sodium acetate 2g, MgSO
47H
2O0.05g, K
2HPO
40.2g, NaCl 0.12g, MnSO
44H
2O 0.01g, FeSO
40.01g, adding distil water is to 1000mL.
Described Nitrate bacteria solid medium is comprised of following component: NaNO
20.5g, sodium acetate 2g, MgSO
47H
2O0.05g, K
2HPO
40.2g, NaCl 0.12g, MnSO
44H
2O 0.01g, FeSO
40.01g, agar 20g, adding distil water be to 1000mL.
Experimental result shows: although be subject to the obvious inhibition of low temperature environment, add a kind of rhodococcus (Rhodococcus sp.) nitre of removing the Nitrite In Polluted Water nitrate nitrogen of the present invention dark-1CGMCC No.5277, at low temperatures, particularly under 8-12 ℃ condition, removal has obvious strengthening effect to the Nitrite In Polluted Water nitrate nitrogen, inoculate rhodococcus of the present invention and compare with the sewage of not inoculating bacterium, the nitrite nitrogen removal efficiency has improved and has been about 10%.
Description of drawings
Fig. 1 is that the low-temperature aerobic condition is gone down except a kind of domestication separation and Culture schema of removing the rhodococcus of Nitrite In Polluted Water nitrate nitrogen of sewage Nitrite Nitrogen.
Fig. 2 is that dark-1 pair of nitrite nitrogen of nitre is removed test.
Embodiment
The present invention is further illustrated below by specific embodiment.
Embodiment 1
The isolation cultivation method of the rhodococcus of removing nitrite nitrogen in sewage under low-temperature condition comprises the steps:
(1) with the sludge seeding of 15mL Tianjin discipline village sewage disposal plant aeration tank to the Erlenmeyer flask that 250mL Nitrate bacteria nutrient solution is housed, nitrogen concentration is 50mg/L in the Sodium Nitrite in the described Nitrate bacteria nutrient solution, at 10 ℃, shaking speed is to tame in the 130rpm shaking table, when treating that the nitrite nitrogen degradation rate reaches 90%, leave standstill, remove supernatant liquor, with the sludge seeding of Erlenmeyer flask bottom to being equipped with in 250mL the second Nitrate bacteria nutrient solution, nitrite nitrogen concentration in described the second Nitrate bacteria nutrient solution is 150mg/L, at 10 ℃, in being the shaking table of 130rpm, rotating speed tames, when treating that the nitrite nitrogen degradation rate reaches 90%, namely obtain containing the Nitrate bacteria nutrient solution of low temperature resistant bacterium;
(2) get the nutrient solution that 10mL step (1) obtains, add oneself and be equipped with in the 250mL Erlenmeyer flask of 90mL sterile purified water and 30 sterilization granulated glass spherees, vibration 20min makes bacterial cell fully be discharged in the supernatant liquor; Leave standstill 30min, get the adding of 1mL supernatant liquor and be equipped with in the test tube of 9mL sterile distilled water, obtain 10
-2Bacteria suspension, take turns doing as stated above serial dilution, accomplish 10 always
-8Till;
(3) choosing gradient is 10
-5～10
-8Bacteria suspension, get respectively 1mL and be inoculated in the Nitrate bacteria solid medium, each extent of dilution is done 3 Duplicate Samples, the numbering; 10 ℃ under the condition of aerobic moisturizing cultivated for 3 weeks;
(4) choosing colony is large in the Nitrate bacteria solid medium flat board from step (3), single bacterium colony that form is different, line separates again in Nitrate 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 nitre is dark-1, the rhodococcus through being accredited as removing nitrite nitrogen in sewage under low-temperature condition (Rhodococcus sp.).
The Nitrate bacteria nutrient solution that adopts among each embodiment is comprised of following component: NaNO
20.25g, sodium acetate 2g, MgSO
47H
2O 0.05g, K
2HPO
40.2g, NaCl 0.12g, MnSO
44H
2O 0.01g, FeSO
40.01g, adding distil water is to 1000mL.
The second Nitrate bacteria nutrient solution is comprised of following component: NaNO
20.74g, sodium acetate 2g, MgSO
47H
2O 0.05g, K
2HPO
40.2g, NaCl 0.12g, MnSO
44H
2O 0.01g, FeSO
40.01g, adding distil water is to 1000mL.
The Nitrate bacteria solid medium is comprised of following component: NaNO
20.5g, sodium acetate 2g, MgSO
47H
2O 0.05g, K
2HPO
40.2g, NaCl 0.12g, MnSO
44H
2O 0.01g, FeSO
40.01g, agar 20g, adding distil water be to 1000mL.
Rhodococcus of the present invention (Rhodococcus sp.) has following Microbiological Characteristics:
1. morphological characteristic
Examine under a microscope, for spherical, diameter is 0.7 μ m; Form the beige bacterium colony at the Nitrate bacteria substratum.
2. cultivate and learn characteristic
Under 10 ℃, when cultivating in the Nitrate bacteria solid medium, Rhodococcus sp. bacterial strain of the present invention has following characteristic.
The surface is more smooth, drying, and the neat in edge rule, individual small rounded, thalline is beige.
3. physiological property
Aerobic condition goes down except the gram positive bacterium of sewage nitrite, catalase is negative, methyl red test is negative, not hydrolyzed starch, liquefy gelatin, produce indoles, growth pH value is not 6～8,5～30 ℃ of growth temperatures.
| Gram | Catalase | Methyl red | Starch Hydrolysis | The gelatin test | Indole test | pH | Temperature |
| G + | - | - | - | - | + | 6～8 | 5～30 |
Learn characteristic according to mentioned microorganism, bacterial strain called after nitre of the present invention dark-1, Classification And Nomenclature is that rhodococcus (Rhodococcus sp) has been preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on September 22nd, 2011, deposit number is CGMCC No.5277, the preservation address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, and survived.
Embodiment 2
The degradation effect of dark-1 pair of nitrite nitrogen of nitre
In the beaker of 1L, add 500mL Nitrate bacteria nutrient solution, initial nitrite nitrogen concentration is adjusted into 50mg/L, inoculate nitre of the present invention dark-1, connecing three rings nitre of the present invention dark-1 from the Nitrate bacteria solid plate enters the above-mentioned Nitrate bacteria nutrient solution, keeping pH is 7.5, measures the degraded situation of 72 hours dark-1 pair of nitrite nitrogens of nitre of the present invention, the results are shown in Figure 2.
As seen from Figure 2, the degraded through 72 hours, nitrogen concentration of nitrite solution is 6.2mg/L, and nitrite nitrogen is removed substantially, and clearance is 87.6%, proves that dark-1 pair of nitrite nitrogen of nitre among the present invention has good removal ability.
Embodiment 3
The isolation cultivation method of the rhodococcus of removing nitrite nitrogen in sewage under low-temperature condition comprises the steps:
(1) with the sludge seeding of 10mL Tianjin discipline village sewage disposal plant aeration tank to the Erlenmeyer flask that 250mL Nitrate bacteria nutrient solution is housed, nitrogen concentration is 50mg/L in the Sodium Nitrite in the described Nitrate bacteria nutrient solution, at 8 ℃, shaking speed is to tame in the 140rpm shaking table, when treating that the nitrite nitrogen degradation rate reaches 80%, leave standstill, remove supernatant liquor, with the sludge seeding of Erlenmeyer flask bottom to being equipped with in 250mL the second Nitrate bacteria nutrient solution, nitrite nitrogen concentration in described the second Nitrate bacteria nutrient solution is 150mg/L, at 8 ℃, in being the shaking table of 140rpm, rotating speed tames, when treating that the nitrite nitrogen degradation rate reaches 80%, namely obtain containing the Nitrate bacteria nutrient solution of low temperature resistant bacterium;
(2) get the nutrient solution that 10mL step (1) obtains, add oneself and be equipped with in the 250mL Erlenmeyer flask of 90mL sterile purified water and 20 sterilization granulated glass spherees, vibration 30min makes bacterial cell fully be discharged in the supernatant liquor; Leave standstill 20min, get the adding of 1mL supernatant liquor and be equipped with in the test tube of 9mL sterile distilled water, obtain 10
-2Bacteria suspension, take turns doing as stated above serial dilution, accomplish 10 always
-8Till;
(3) choosing gradient is 10
-5～10
-8Bacteria suspension, get respectively 1mL and be inoculated in the Nitrate bacteria solid medium, each extent of dilution is done 3 Duplicate Samples, the numbering; 10 ℃ under the condition of aerobic moisturizing cultivated for 2 weeks;
(4) choosing colony is large in the Nitrate bacteria solid medium flat board from step (3), single bacterium colony that form is different, line separates again in Nitrate bacteria solid medium flat board, repeating plate streaking separates 4 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 nitre is dark-1, the rhodococcus through being accredited as removing nitrite nitrogen in sewage under low-temperature condition (Rhodococcus sp.).
Embodiment 4
The isolation cultivation method of the rhodococcus of removing nitrite nitrogen in sewage under low-temperature condition comprises the steps:
(1) with the sludge seeding of 20mL Tianjin discipline village sewage disposal plant aeration tank to the Erlenmeyer flask that 250mL Nitrate bacteria nutrient solution is housed, nitrogen concentration is 50mg/L in the Sodium Nitrite in the described Nitrate bacteria nutrient solution, at 12 ℃, shaking speed is to tame in the 120rpm shaking table, when treating that the nitrite nitrogen degradation rate reaches 90%, leave standstill, remove supernatant liquor, with the sludge seeding of Erlenmeyer flask bottom to being equipped with in 250mL the second Nitrate bacteria nutrient solution, nitrite nitrogen concentration in described the second Nitrate bacteria nutrient solution is 150mg/L, at 12 ℃, in being the shaking table of 120rpm, rotating speed tames, when treating that the nitrite nitrogen degradation rate reaches 90%, namely obtain containing the Nitrate bacteria nutrient solution of low temperature resistant bacterium;
(2) get the nutrient solution that 10mL step (1) obtains, add oneself and be equipped with in the 250mL Erlenmeyer flask of 90mL sterile purified water and 30 sterilization granulated glass spherees, vibration 10min makes bacterial cell fully be discharged in the supernatant liquor; Leave standstill 40min, get the adding of 1mL supernatant liquor and be equipped with in the test tube of 9mL sterile distilled water, obtain 10
-2Bacteria suspension, take turns doing as stated above serial dilution, accomplish 10 always
-8Till;
(3) choosing gradient is 10
-5～10
-8Bacteria suspension, get respectively 1mL and be inoculated in the Nitrate bacteria solid medium, each extent of dilution is done 3 Duplicate Samples, the numbering; 10 ℃ under the condition of aerobic moisturizing cultivated for 3 weeks;
(4) choosing colony is large in the Nitrate bacteria solid medium flat board from step (3), single bacterium colony that form is different, line separates again in Nitrate 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 nitre is dark-1, goes down except the rhodococcus (Rhodococcus sp.) of sewage nitrite chlorine through being accredited as cold condition.
Experiment showed, that embodiment 3 and embodiment 4 separation and Culture remove the effect that the rhodococcus (Rhodococcus sp.) of Nitrite In Polluted Water nitrate nitrogen also has good degrading nitrite nitrogen, clearance is respectively 87.0% and 87.2%.
Claims (2)
1. dark-1 CGMCCNo.5277 of the rhodococcus of a removing nitrite nitrogen in sewage under low-temperature condition (Rhodococcus sp.) nitre, it has in the low-temperature aerobic condition to go down except the ability of sewage Nitrite Nitrogen, and described low temperature is 8～12 ℃.
2. dark-1 CGMCC No.5277 of the rhodococcus of a kind of removing nitrite nitrogen in sewage under low-temperature condition of claim 1 (Rhodococcus sp.) nitre goes down except the purposes of sewage Nitrite Nitrogen in the low-temperature aerobic condition, and described low temperature is 8～12 ℃.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001212594A (en) * | 2000-02-04 | 2001-08-07 | Kansai Paint Co Ltd | Method for removing nitrate nitrogen in wastewater |
| CN101875909A (en) * | 2010-01-26 | 2010-11-03 | 中国科学院成都生物研究所 | Heterotrophic nitrification aerobic denitrifying bacteria and culture method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001212594A (en) * | 2000-02-04 | 2001-08-07 | Kansai Paint Co Ltd | Method for removing nitrate nitrogen in wastewater |
| CN101875909A (en) * | 2010-01-26 | 2010-11-03 | 中国科学院成都生物研究所 | Heterotrophic nitrification aerobic denitrifying bacteria and culture method and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| 一株轻度嗜盐反硝化菌的分离鉴定及特性;郭艳丽;《应用与环境生物学报》;20100625;第16卷(第3期);394-398 * |
| 郭艳丽.一株轻度嗜盐反硝化菌的分离鉴定及特性.《应用与环境生物学报》.2010,第16卷(第3期), |
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Inventor after: Ma Baoling Inventor after: Tian Qing Inventor after: Hao Bin Inventor after: Chen Aiyin Inventor after: Jiang Xiuguang Inventor after: Fan Qian Inventor after: Bao Xiaobo Inventor after: Wu Duan Inventor before: Tian Qing Inventor before: Jiang Xiuguang Inventor before: Hao Bin Inventor before: Chen Aiyin |
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Free format text: CORRECT: INVENTOR; FROM: TIAN QING JIANG XIUGUANG HAO YUN CHEN AIYIN TO: MA BAOLING TIAN QING HAO YUN CHEN AIYIN JIANG XIUGUANG FAN QIAN BAO XIAOBO WU DUAN |
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Effective date of registration: 20201231 Address after: Room 405-406, floor 4, w5-c2-51, Third Street, Tianjin Economic and Technological Development Zone, Binhai New Area, Tianjin Patentee after: CAPITAL AIHUA (TIANJIN) MUNICIPAL & ENVIRONMENTAL ENGINEERING Co.,Ltd. Address before: Room 402, 44 shuishanggongyuan West Road, Nankai District, Tianjin Patentee before: WATER TREATMENT NEW TECHNOLOGY INDUSTRIALIZATION BASE, MINISTRY OF CONSTRUCTION (TIANJIN PORT FREE TRADE ZONE WATER TREATMENT NEW TECHNOLOGY INDUSTRIALIZATION BASE) |