CN104140935A - Denitrification rhodococcus and rhodococcus microbial agent production method and application - Google Patents
Denitrification rhodococcus and rhodococcus microbial agent production method and application Download PDFInfo
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- CN104140935A CN104140935A CN201310165610.2A CN201310165610A CN104140935A CN 104140935 A CN104140935 A CN 104140935A CN 201310165610 A CN201310165610 A CN 201310165610A CN 104140935 A CN104140935 A CN 104140935A
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Abstract
The invention relates to rhodococcus with the function of removal of ammonia nitrogen and nitrite in water and a low cost rhodococcus microbial agent production method and an application method, and belongs to the technical field of environmental microorganisms. The adopted strain is an efficient denitrification bacteria-(Rhodococcus sp.) YY12 strain separated and screened from activated sludge of sewage treatment plants, and the strain is preserved in China General Microbiological Culture Collection Center (CGMCC) with the preservation number of CGMCC No. 6924. According to the method, monosodium glutamate wastewater is used as a fermentation medium for large-scale culture of the Rhodococcus, the microbial agent for efficient removal of ammonia nitrogen and nitrite in water is prepared, and the process has the advantages of low production cost, simple operation, resource utilization of the monosodium glutamate wastewater, and reduction of pollution emissions. The microbial agent used for aquaculture not only can improve the water quality of aquaculture water, but also can significantly improve the aquatic products output, and shows a wide application prospect.
Description
Technical field:
The present invention relates to a strain and there is the rhodococcus of removing ammonia nitrogen and nitrite function in water body, and low cost production method and the using method of microbial inoculum, environmental microorganism technical field belonged to.
Background technology:
Ammonia-nitrogen content is too high is one of principal element of water pollution and eutrophication.Microbial denitrogenation method is extensively approved with all advantages such as its high-level efficiency, low cost, non-secondary pollution, becomes the focus that environmental science and water treatment field are paid close attention to research, but traditional wastewater biological denitrificaion technique still exists problems.The appearance of synchronous nitration and denitrification bio-denitrification technology in recent years has not only overcome the problem that traditional biological denitrogenation exists, but also there is advantages such as shortening denitrogenation course, saving carbon source, reduction energy consumption, the design that improves processing power, simplified system and operation, there is very large development potentiality.
Heterotrophic nitrification refer to microorganism when utilizing organic substrates by mineralized nitrogen the process for azanol, nitrite and nitrate.Aerobic denitrification refers to that microorganism can utilize oxygen and nitrate or nitrite to carry out respiration for electron acceptor(EA) simultaneously
[17].The discovery of nitrification bacteria and aerobic denitrifying bacteria etc., broken traditional theory and thought that nitration reaction can only complete the understanding that can only carry out at anaerobic condition with anti-nitration reaction by autotrophic bacteria, make synchronous nitration and denitrification (Simultaneous Nitrification and Denitrification, SND) become possibility.Inventor herein separates a strain from the active sludge of sewage work can have heterotrophic nitrification-aerobic denitrification function denitrifying bacteria simultaneously, for the removal of ammonia nitrogen in sewage and the better popularization of probiotics are laid a good foundation.
In recent years, the research that microbiobacterial agent purifies water and application are more and more deeply with extensive.Chinese invention patent (application number: disclose a strain rhodococcus (Rhodococcus sp.) BZ-9 bacterial strain 201010156031.8), deposit number is CGMCC No. 3695, multiple toxic organic compound matter is had to efficient degradation performance, and its microbial inoculum is applicable to the processing of the organic waste water that contains the toxic organic compound matter such as phenol and pyridine that produces in coking chemical waste water, leather-making waste water, dyeing waste water and other petrochemical production process; Its fermentation culture based component is: glucose 0.8%, (NH
4)
2sO
41%, K
2hPO
40.2%, MgSO
40.05%, NaCl 0.01%, CaCO
30.3%, yeast extract paste 0.02%, and all the other are water, pH value 7.2-7.5.Chinese invention patent (application number: disclose a strain 201110006790.0) and had a liking for pyridine rhodococcus (Rhodococcus pyridinivorans) CPZ-24 bacterial strain, deposit number is CGMCC No. 4401, this bacterial strain has heterotrophic nitrification and aerobic denitrification, this bacterium is in denitrification process, and the gas of generation 99% is N
2, also have a small amount of N
2o, NO, NO
2; The substratum of this bacterial strain mainly provides carbon source with sodium succinate, (NH
4)
2sO
4nitrogenous source is provided, also has K
2hPO
4, NaCl, MgSO
4, MnSO
4, FeSO
4in inorganic salt, form.Chinese invention patent (application number: disclose a strain Rhodococcus strain (nitre dark-1) 201110416893.4), deposit number is CGMCC No. 5277,5277, and it has in low-temperature aerobic condition to go down except the ability of sewage Nitrite Nitrogen.Particularly under the condition of 8-12 ℃, Nitrite In Polluted Water nitrate nitrogen is removed and had obvious strengthening effect, to inoculate rhodococcus of the present invention and compare with the sewage of not inoculating bacterium, nitrite nitrogen removal efficiency has improved and has been about 10%; The substratum of this bacterial strain with sodium acetate for providing carbon source, (NH
4)
2sO
4nitrogenous source is provided, also has K
2hPO
4, NaCl, MgSO
4, MnSO
4, FeSO
4in inorganic salt, form.
In sum, the bacterial strain report with denitrification functions is more, and its cultural method is also different, still, has no and adopts gourmet powder waste water to cultivate the report that has denitrification functions rhodococcus and prepare microbial inoculum.Gourmet powder waste water has high COD, high BOD, high sulfate radical content, high ammonia-nitrogen content, feature that biodegradability is good mostly.The wastewater flow rate producing in glutamate production process is very large, more difficult to the processing of these waste water.It is reported, every production 1 t monosodium glutamate, approximately will discharge 10～15 t and extract the mother liquor after L-glutamic acid, and the whole nation will be discharged more than 1,000 ten thousand tons of this high concentrated organic wastewaters every year.These wastewater treatment qualified discharge costs are high, the method for urgently seeking recycling.
Summary of the invention:
The object of this invention is to provide a strain and there is denitrification functions rhodococcus, and a kind of method of can recycling monosodium glutamate powder waste water cultivating rhodococcus and preparing microbial inoculum is provided, and utilize this microbial inoculum to remove the using method of ammonia nitrogen and nitrite in aquaculture water.
Bacterial strain used in the present invention is by contriver's separation screening plant height effect out from the active sludge of sewage work, to be removed rhodococcus (Rhodococcus sp.) the YY12 bacterial strain of ammonia nitrogen in sewage and nitrite, this bacterial strain is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center on December 3rd, 2012, deposit number is: CGMCC No. 6924, depositary institution address: Datun Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode: 100101.
Production method and the using method of rhodococcus denitrification microorganism microbial inoculum of the present invention are as follows:
1, gourmet powder waste water pre-treatment: regulate pH to 6.8～8.0 of waste water with aqueous sodium hydroxide solution or liming, in 110～126 ℃ of sterilizings 10～35 minutes.
2, liquid seeds preparation: to the rhodococcus of inoculating slant preservation in the triangular flask of the gourmet powder waste water that pre-treatment is good is housed, in 25～35 ℃, 120～200 revs/min of shaking culture 24～36 hours, obtain liquid seeds.
3, liquid state fermentation: to inoculate volume ratio in the gourmet powder waste water fermentor tank that pre-treatment is good be 2%～15% liquid seeds to being equipped with, and in 25～35 ℃, logical sterile air is cultivated 15～36 hours; Cultured fermented liquid is through count plate, and viable count reaches 1.0 * 10
9more than CFU/ milliliter, gained fermented liquid can be used as liquid bacterial agent and directly uses.
4, solid fungicide preparation: the zeolite of 80 mesh sieves excessively of take is carrier, according to fermented liquid: zeolite is the weight ratio of 1: 2～1: 10, fermented liquid is added in zeolite, fully mix, obtain solid-state microbial inoculum, air seasoning, is packed below 20% to microbial inoculum water content, leave shady and cool dry place in, standby.
The using method of rhodococcus microbiobacterial agent 5: by fermented liquid or 10-100 times of clear water dilution for solid fungicide, mix, consumption according to every cubic metre of water body 10-100 milliliter diluent is evenly splashed in the pool of aquaculture, splash 1 5-10 days summer, and splash 1 10-20 days other seasons.
Embodiment:
By the using method of rhodococcus provided by the invention and microbial inoculum thereof, can remove fast ammonia nitrogen and nitrite in aquaculture water, improve aquaculture water environment.Embodiment is as follows:
Embodiment 1: gourmet powder waste water is produced rhodococcus microbiobacterial agent
Gourmet powder waste water pre-treatment: regulate wastewater pH to 7.5 with liming, in 115 ℃ of sterilizings 20 minutes.
Liquid seeds preparation: to the rhodococcus of inoculating slant preservation in the triangular flask of the gourmet powder waste water that pre-treatment is good is housed, in 30 ℃, 150 revs/min of shaking culture 30 hours, obtain liquid seeds.
Liquid state fermentation: inoculate 5%(volume ratio to being equipped with in the gourmet powder waste water fermentor tank that pre-treatment is good) liquid seeds, in 32 ℃, logical sterile air is cultivated 30 hours; Cultured fermented liquid is through count plate, and viable count reaches 2.0 * 10
9cFU/ milliliter.
Microbial inoculum preparation: the zeolite of crossing 80 mesh sieves of take is carrier, according to fermented liquid: the weight ratio that zeolite is 1: 3, fermented liquid is added in biological carbon, fully mix, obtain solid-state microbial inoculum, air seasoning is to microbial inoculum water content below 20%, packing, leaves shady and cool dry place in.
Embodiment 2: rhodococcus microbiobacterial agent is applied to improve aquaculture water quality
Experimental Base is positioned at Coastal Area of Guangdong Province, by the high-order cultivating pool of 4 Penaeus vannameis, (each pond is taken up an area 3 mu, dark 2.5 meters of pond) set up two the contrast pool and 2 processing pools that add microbial inoculum that do not add microbial inoculum separately, shrimp seedling 360,000 tails are thrown in each pond, and breeding way is identical.By 20 times of clear water dilutions for solid fungicide, mix, according to the consumption of 50 milliliters of diluents of every cubic metre of water body, evenly to splash to two and process in the pools, splash 17 days summers, and splash 1 15 days other seasons.The aquaculture water ammonia-nitrogen content of processing the pool can be stablized and is controlled at below 0.5 mg/litre, and nitrite nitrogen can be controlled in below 0.8 mg/litre; And aquaculture water ammonia-nitrogen content fluctuates between 0.5～1.5 mg/litre in the contrast pool, and temperature is higher, and ammonia nitrogen concentration is larger, and nitrite nitrogen can fluctuate the content most of the time between 1.0～2.5 mg/litre, along with temperature raises and raises.Results when Penaeus vannamei specification reaches 40-50 tail/jin, improve 10% left and right in the yield increased pool of prawn in the processing pool.
Claims (3)
1. a strain has the rhodococcus (Rhodococcus sp.) of removing ammonia nitrogen and nitrite function in water body, it is characterized in that: this strain rhodococcus is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number: CGMCC No. 6924.
2. rhodococcus claimed in claim 1 is produced the method for denitrification microorganism microbial inoculum, comprises the steps:
(1) gourmet powder waste water pre-treatment: regulate pH to 6.8～8.0 of waste water with aqueous sodium hydroxide solution or liming, in 110～126 ℃ of sterilizings 10～35 minutes;
(2) liquid seeds preparation: to the rhodococcus of inoculating slant preservation in the triangular flask of the gourmet powder waste water that pre-treatment is good is housed, in 25～35 ℃, 120～200 revs/min of shaking culture 24～36 hours, obtain liquid seeds;
(3) liquid state fermentation: to inoculate volume ratio in the gourmet powder waste water fermentor tank that pre-treatment is good be 2%～15% liquid seeds to being equipped with, and in 25～35 ℃, logical sterile air is cultivated 15～36 hours; Cultured fermented liquid is through count plate, and viable count reaches 1.0 * 10
9more than CFU/ milliliter, gained fermented liquid can be used as liquid bacterial agent and directly uses;
(4) solid fungicide preparation: the zeolite of 80 mesh sieves excessively of take is carrier, according to fermented liquid: zeolite is the weight ratio of 1: 2～1: 10, fermented liquid is added in zeolite, fully mix, obtain solid-state microbial inoculum, air seasoning, is packed below 20% to microbial inoculum water content, leave shady and cool dry place in, standby.
3. the using method of denitrification microorganism microbial inoculum claimed in claim 2, it is characterized in that: by fermented liquid or 10-100 times of clear water dilution for solid fungicide, mix, consumption according to every cubic metre of water body 10-100 milliliter diluent is evenly splashed in the pool of aquaculture, splash 1 5-10 days summer, and splash 1 10-20 days other seasons.
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|---|---|---|---|
| CN201310165610.2A CN104140935B (en) | 2013-05-08 | 2013-05-08 | One plant of production method and application of the Rhodococcus sp and its microbial inoculum with denitrification functions |
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| CN201310165610.2A CN104140935B (en) | 2013-05-08 | 2013-05-08 | One plant of production method and application of the Rhodococcus sp and its microbial inoculum with denitrification functions |
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104611279A (en) * | 2015-03-02 | 2015-05-13 | 中蓝连海设计研究院 | Rhodococcus erythropolis LH-N13 as well as microbial agent and use thereof |
| CN104671416A (en) * | 2015-02-15 | 2015-06-03 | 中国科学院生态环境研究中心 | Device for restoring eutrophic water |
| CN105039228A (en) * | 2015-09-05 | 2015-11-11 | 内蒙古阜丰生物科技有限公司 | Biological agent for glutamate wastewater treatment |
| CN105152478A (en) * | 2015-09-09 | 2015-12-16 | 内蒙古阜丰生物科技有限公司 | Treatment method of industrial wastewater produced during preparation of sodium glutamate through concentration isoelectric process |
| CN105174443A (en) * | 2015-09-09 | 2015-12-23 | 呼伦贝尔东北阜丰生物科技有限公司 | Treatment process for wastewater produced by extracting monosidum glutamate by concentrating isoelectric points |
| CN107400645A (en) * | 2017-07-31 | 2017-11-28 | 河北科技大学 | Composite bacteria agent for repairing polluted by nitrogen soil and preparation method thereof |
| CN107828679A (en) * | 2017-10-27 | 2018-03-23 | 中国水产科学研究院南海水产研究所 | The rose-red Rhodococcus strain XHRR1 of ammonia and its application in a kind of purifying aquatic water |
| CN108522381A (en) * | 2018-04-19 | 2018-09-14 | 广州普麟生物制品有限公司 | A kind of closed zero-emission batch production culture of Penaeus vannamei method |
| CN111099743A (en) * | 2018-10-26 | 2020-05-05 | 中国石油化工股份有限公司 | Composition for enhancing denitrification and denitrification of wastewater and denitrification method thereof |
| CN112574921A (en) * | 2020-12-26 | 2021-03-30 | 北京华晨吉光科技有限公司 | Method for preparing aerobic denitrification composite microbial inoculum by utilizing kitchen waste and application thereof |
| CN114292782A (en) * | 2021-12-28 | 2022-04-08 | 江苏南资环保科技有限公司 | Bacterium for strengthening facultative FMBR pesticide wastewater treatment process |
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| CN104671416A (en) * | 2015-02-15 | 2015-06-03 | 中国科学院生态环境研究中心 | Device for restoring eutrophic water |
| CN104671416B (en) * | 2015-02-15 | 2016-12-07 | 中国科学院生态环境研究中心 | Repair the device of eutrophication water |
| CN104611279A (en) * | 2015-03-02 | 2015-05-13 | 中蓝连海设计研究院 | Rhodococcus erythropolis LH-N13 as well as microbial agent and use thereof |
| CN104611279B (en) * | 2015-03-02 | 2017-06-09 | 中蓝连海设计研究院 | A kind of red city Rhodococcus sp LH N13 and its microbial bacterial agent and purposes |
| CN105039228A (en) * | 2015-09-05 | 2015-11-11 | 内蒙古阜丰生物科技有限公司 | Biological agent for glutamate wastewater treatment |
| CN105152478A (en) * | 2015-09-09 | 2015-12-16 | 内蒙古阜丰生物科技有限公司 | Treatment method of industrial wastewater produced during preparation of sodium glutamate through concentration isoelectric process |
| CN105174443A (en) * | 2015-09-09 | 2015-12-23 | 呼伦贝尔东北阜丰生物科技有限公司 | Treatment process for wastewater produced by extracting monosidum glutamate by concentrating isoelectric points |
| CN107400645A (en) * | 2017-07-31 | 2017-11-28 | 河北科技大学 | Composite bacteria agent for repairing polluted by nitrogen soil and preparation method thereof |
| CN107828679A (en) * | 2017-10-27 | 2018-03-23 | 中国水产科学研究院南海水产研究所 | The rose-red Rhodococcus strain XHRR1 of ammonia and its application in a kind of purifying aquatic water |
| CN107828679B (en) * | 2017-10-27 | 2021-04-27 | 中国水产科学研究院南海水产研究所 | Rhodococcus roseus strain XHRR1 for purifying ammonia in aquaculture water and application thereof |
| CN108522381A (en) * | 2018-04-19 | 2018-09-14 | 广州普麟生物制品有限公司 | A kind of closed zero-emission batch production culture of Penaeus vannamei method |
| CN111099743A (en) * | 2018-10-26 | 2020-05-05 | 中国石油化工股份有限公司 | Composition for enhancing denitrification and denitrification of wastewater and denitrification method thereof |
| CN111099743B (en) * | 2018-10-26 | 2022-06-07 | 中国石油化工股份有限公司 | Composition for enhancing denitrification and denitrification of wastewater and denitrification method thereof |
| CN112574921A (en) * | 2020-12-26 | 2021-03-30 | 北京华晨吉光科技有限公司 | Method for preparing aerobic denitrification composite microbial inoculum by utilizing kitchen waste and application thereof |
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