CN111621438A - Wedner mannich bacillus LM-LZ separated from oxidation pond of pig farm and application thereof - Google Patents
Wedner mannich bacillus LM-LZ separated from oxidation pond of pig farm and application thereof Download PDFInfo
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Abstract
The invention discloses a Weid Mannich Bacillus (Bacillus Wiedmannii) LM-LZ separated from bottom mud of a pig farm oxidation pond, which is the Weid Mannich Bacillus Wiedmannii with the preservation number: CGMCC No. 18768. The invention also discloses the application of the strain: treating the nitrogen-containing wastewater. The strain belongs to heterotrophic nitrification-aerobic denitrification bacteria, and can effectively remove total nitrogen in wastewater.
Description
Technical Field
The invention relates to a heterotrophic nitrification-aerobic denitrification strain widmanmannii (Bacillus Wiedmannii) LM-LZ separated from bottom mud of a biogas slurry oxidation pond and application thereof in treatment of nitrogen-containing wastewater.
Background
With the social development and the intensification of the production activity of people, a large amount of wastewater containing pollutants is discharged into natural water without treatment, so that the eutrophication of water bodies and a series of environmental problems are increasingly prominent. The nitrogen is an important nutritional pollutant formed by water eutrophication, and how to treat and remove the nitrogen in the wastewater has great significance for eliminating the water eutrophication caused by the nitrogen and protecting the water environment.
Nitrogen is a ubiquitous pollutant in sewage, nitrogen in a water body exists in the form of organic nitrogen and inorganic nitrogen, specific forms of the nitrogen include protein nitrogen, ammonia nitrogen, organic nitrogen, nitrite nitrogen and nitrate nitrogen, and how to efficiently and quickly remove the nitrogen in water is an important subject and a hotspot in the field of current water pollution prevention and treatment technology research. The denitrification process is considered to be a strict anaerobic process by the traditional theory, but since the 80 th generation of the 20 th century, aerobic denitrifying bacteria are continuously reported, and researchers begin to concern about the denitrification potential and the action mechanism of the aerobic denitrifying bacteria. Researches find that the aerobic denitrifying bacteria contain an aerobic denitrifying enzyme system, which comprises two nitrate reductases existing in bacterial cells, namely, nitrate reductase in the membrane and periplasmic nitrate reductase combined with the membrane, wherein the nitrate reductase in the membrane does not work when being aerobic, and the periplasmic nitrate reductase can still work under the condition of high dissolved oxygen concentration; nitrite reductase distributed in the periplasm of cell membrane, including one of cytochrome cd1 type nitrite reductase and the other copper-containing ionic type nitrite reductase; nitric oxide reductase, bound to cell membrane, catalyzes the reduction of NO to N2O; a nitrous oxide reductase, which is a soluble copper ion-containing enzyme, and which is localized in the periplasmic space of the membrane2Reduction of O to N2. The synergistic respiration is an important mechanism of aerobic denitrification, namely, the denitrifying bacteria can transfer electrons from reduced substances to oxygen and simultaneously transfer the electrons to nitrate. The traditional biological denitrification technology is based on nitrification and denitrification of microorganisms, has more defects, has the problems of discontinuity and low efficiency in the denitrification process, finds aerobic denitrifying bacteria, and establishes a novel sewage denitrification technology for upgrading the sewage denitrification treatment technology and overcoming the defects of the traditional biological denitrification technologyThe denitrification technology provides theoretical support.
Other bacillus species are mainly known as closely related microorganisms for denitrification treatment of sewage, and some other bacillus species have been studied to show strong denitrification (plum, forest iron, journal of aquatic ecology, 2009, 2 (3): 48-52; guo, Master thesis of university of tai-rationale 2015; Fan LF, Shieh WY, Wu WF, et al, 2006, coast and Shelf Science,69: 543-. As for the research of the Wedner Mannich bacillus on the denitrification potential of sewage, the research is not reported at home and abroad.
Disclosure of Invention
The invention aims to solve the technical problem of providing a heterotrophic nitrification-aerobic denitrification strain widmanmannii (Bacillus Wiedmannii) LM-LZ for treating nitrogen-containing wastewater and application thereof.
In order to solve the technical problems, the invention provides a heterotrophic nitrification-aerobic denitrification strain widmanmannii (Bacillus Wiedmannii) LM-LZ separated from bottom mud of a pig farm oxidation pond (live pig farm biogas slurry oxidation pond bottom mud), which is widmanmannii Bacillus Wiedmannii with a preservation number: CGMCC No. 18768.
As an improvement of the Wedner Mannich Bacillus (Bacillus Wiedmannii) LM-LZ of the invention: the 16S rDNA gene sequence of the strain is shown in SEQ ID NO. 1.
The invention also provides the application of the Weide Mannich Bacillus (Bacillus Wiedmannii) LM-LZ: nitrogen-containing wastewater (nitrogen-rich wastewater) is treated.
As an improvement of the use of the Wedner Mannich Bacillus (Bacillus Wiedmannii) LM-LZ of the invention: removing nitrogen in the wastewater; belongs to heterotrophic nitrification-aerobic denitrifying bacteria, and can effectively remove the total nitrogen in the wastewater.
The preservation information of the strains of the invention is as follows:
the preservation name is: wideher mannich Bacillus Wiedmannii, depository: china general microbiological culture Collection center, preservation Address: the microbial research institute of the national academy of sciences No. 3, Xilu No.1, Beijing, Chaoyang, Beijing, with the preservation number: CGMCC No.18768, preservation date: 2019, 10 and 30.
The bacterial colony characteristics of the strain are as follows: the bacillus brevis is a bacillus brevis, has rough surface, pilus and no cytopenia, has small capsule and gram-positive bacteria, has the size of (1.2-1.7) Mumx (0.4-0.7) Mum, and has circular bacterial colony, convex and smooth surface, complete edge, white color and opacity on a solid culture medium.
The 16S rDNA gene sequence of the strain is shown in SEQ ID NO. 1:
the strain is obtained by screening biogas slurry oxidation pond bottom sludge collected from a water north town live pig farm in Yushu district of Xinyu province, and according to a fatty acid map generated by qualitative and quantitative analysis of a Sherlock MIS software system on a strain LM-LZ, comparing with a Library database, preliminarily identifying that the strain LM-LZ is Wedner Mannich Bacillus (Bacillus Wiedmannii), and the similarity index SI (similarity index) is 0.991; the strain is also proved to be Wedney Mannich Bacillus (Bacillus Wiedimanii) by a 16S rDNA sequencing identification method, and the similarity index SI (similarity index) is 100.
Inoculating the strain of the invention into nitrogen-containing wastewater, and carrying out fermentation at 15-35 ℃ and 150-200 r.min-1The culture is carried out for 2-3 d under the aerobic condition, and the total nitrogen, ammonia nitrogen, nitrate nitrogen and nitrite nitrogen in the wastewater can be effectively removed.
The total nitrogen concentration of the nitrogen-containing wastewater is 60-600 mg.L-1The ammonia nitrogen concentration is 20-200 mg.L-1The total phosphorus concentration of nitrate nitrogen is 20-250 mg.L-1The ammonia nitrogen concentration of nitrite nitrogen is 20-200 mg.L-1。
The strain can be used for treating pig breeding wastewater, and the pH value is 7.0, the temperature is 25-30 ℃, and the OD is OD415nm0.2, the total nitrogen in the nitrogen-containing wastewater is 593.0 mg.L-1194.8 mg.L of ammonia nitrogen-1The nitrate nitrogen is 202.2 mg.L-1And nitrite nitrogen of 196.0 mg.L-1At a rotation speed of 180 r.min-1Culturing for 3 days, filtering, and collecting the total nitrogen, ammonia nitrogen and water in the wastewater,The removal rates of nitrate nitrogen and nitrite nitrogen are respectively 96.71%, 97.42%, 95.93% and 96.78%, and can reach the first-grade B limit standard (the total nitrogen concentration is 20 mg.L) of pollutant emission standard (GB18918-2002) of urban sewage treatment plant-1The ammonia nitrogen concentration is 8 mg.L-1) And (5) discharging.
The heterotrophic nitrification-aerobic denitrification strain Wedney Mannich Bacillus (Bacillus Wiedmannii) LM-LZ and the application thereof in the treatment of nitrogen-containing wastewater belong to the discovery for the first time.
In conclusion, the heterotrophic nitrification-aerobic denitrification strain widmannhi (Bacillus Wiedmannii) LM-LZ for treating the nitrogen-containing wastewater is obtained by screening, and has the potential of efficiently treating the nitrogen-containing wastewater. Provides a technical basis for controlling the influence of the discharge of the nitrogenous wastewater on the environment, and has wide application prospect.
Drawings
The following describes embodiments of the present invention in further detail with reference to the accompanying drawings.
FIG. 1 shows Wedner Mannich Bacillus (Bacillus Wiedmannii) LM-LZ (magnified 10000 times);
FIG. 2 shows the result of 16S rDNA PCR amplification (LZ: strain LM-LZ; Marker: DNA standard molecular weight);
FIG. 3 is a phylogenetic tree of strain LM-LZ based on the 16S rDNA sequence.
Detailed Description
The invention will be further described with reference to specific examples, but the scope of the invention is not limited thereto:
example 1 screening and identification of degrading Strain
1 materials and methods
1.1 media and reagents
Enrichment culture medium: potassium nitrate (2, 5, 10, 20 or 30g, gradually increased and added for acclimatization and enrichment), CaCl2·6H2O0.1g，MgCl20.25g，K2HPO41.5g，NH4Cl 1g, peptone 9g, H2O 1000mL，pH＝6.5～7.0；
Basic culture medium: NH (NH)4NO31.00g，MgSO4·7H2O 0.5g，(NH4)2SO40.5g，KH2PO40.5g，NaCl0.5g，K2HPO41.5g，H2O 1000mL，pH4.0；
Adding 30g potassium nitrate into each 1000ml basal medium to obtain a medium containing 30 g.L-1A liquid basal medium of potassium nitrate;
adding 30g potassium nitrate and 15g agar into 1000ml basal medium to obtain culture medium containing 30 g.L-1A solid basal medium of potassium nitrate;
all media need to be sterilized at high temperature before use (conventional high temperature sterilization, generally sterilization at 1.1 atm and 121 ℃ for 20min), which is common knowledge.
Nitrogen-containing wastewater: from 20.48 g.L-1Sodium acetate, 1.44 g.L-1Potassium nitrate, 0.98 g.L-1Sodium nitrite, 0.94 g.L-1Ammonium sulfate, 0.5 g.L-1K2HPO4·3H2O and 0.03 g.L-1MgSO4·7H2O, pH 7.0. Wherein the total nitrogen is 593.0 mg.L-1194.8 mg. L of ammonia nitrogen-1202.2 mg.L of nitrate nitrogen-1196.0 mg.L of nitrite nitrogen-1。
1.2 determination of the effectiveness of the Strain in the treatment of Nitrogen-containing wastewater
Propagating the purified single purified strain through a basic culture medium, and specifically comprising the following steps: inoculating one loop of the purified strain to 100mL of basic foundation at 30 ℃ for 180 r.min-1Culturing on a shaking table for 7 days; the obtained culture solution is transferred into 1000mL of liquid basal culture medium according to the inoculation amount of 10 percent (volume percent), and the culture is continued for 3 days under the same culture conditions; and obtaining the propagation bacterium liquid. Measured as bacterial load OD415nmThe resulting mixture was inoculated into a container containing 100mL of the above nitrogen-containing wastewater (20.48 g. L.)-1Sodium acetate, 1.44 g.L-1Potassium nitrate, 0.98 g.L-1Sodium nitrite, 0.94 g.L-1Ammonium sulfate, 0.5 g.L-1K2HPO4·3H2O and 0.03 g.L-1MgSO4·7H2O, pH7.0)250mL of triangular flask, namely inoculating the propagation medium (about 10mL) into 100mL of nitrogen-containing wastewaterIn the 250mL triangular flask of (1), the bacterial count OD415nm in the wastewater at this time was 0.2; using nitrogen-containing wastewater without inoculation as control, at 30 deg.C and 180 r.min-1The aerobic culture is carried out for 3 days on a constant temperature shaking table. After the culture, the mycelium was removed by filtration through a 0.20 μm membrane, and the filtrate was used for measurement of total nitrogen, ammonia nitrogen, nitrate nitrogen and nitrite nitrogen (see: the institute of Water and wastewater monitoring and analysis methods of the State environmental protection Bureau, ed., Japan, 2002, published by the national environmental sciences).
Removal rate (%) × 100 (control sample residual amount-treated sample residual amount) × 100/control sample residual amount.
1.3 dominant degradation Strain Breeding
1.3.1 sources of strains
Collecting biogas slurry oxidation pond sediment of a water north town live pig farm in Yu water areas of Xinyu province in Jiangxi province.
1.3.2 isolation, purification and screening of degrading strains
Taking 10g of bottom sludge from the biogas slurry oxidation pond of each pig farm, respectively adding the bottom sludge into a 250mL triangular flask filled with 100mL of sterile liquid enrichment medium under the sterile condition, and carrying out 180 r-min at 30 DEG C-1After culturing for 7 days on a shaking table, the cells were transferred to the next enrichment medium according to the inoculum size of 10% (potassium nitrate gradient is 2, 5, 10, 20 or 30 g.L in sequence)-1) And domesticating and culturing for 7 days under the same condition. By analogy, the potassium nitrate concentration is 30 g.L-1And (5) enriching culture solution corresponding to the culture medium.
The culture solution is transferred to a medium containing 30 g.L according to the inoculation amount of 10 percent-1Continuously culturing in liquid basic culture medium containing potassium nitrate for 7 days, and continuously inoculating for 2 times; taking 0.1mL of the obtained fermentation broth to a fermentation broth content of 30 g.L-1Repeatedly carrying out plate streaking separation and purification on a solid basic culture medium of potassium nitrate until a single bacterial colony is obtained by screening, inoculating the pure bacterial colony on a slope, and storing in a refrigerator at 4 ℃.
The strains obtained by screening are detected according to the method for measuring the treatment efficiency of the nitrogen-containing wastewater reported above, and the strain with the best treatment effect on the nitrogen-containing wastewater is selected as the strain of the invention.
1.3.3 Strain identification
The strain identification adopts a Sherlock Microbiological Identity System (MIS) software system of MIDI company in America, and the system extracts and analyzes fatty acid of a single colony purified on a basic culture medium according to the operation specification of the MIDI company, qualitatively (type) and quantitatively (content) analyzes the fatty acid component of a microorganism and generates a fatty acid map, the generated map is compared with a database (Library), unknown strains are identified according to a similarity index SI (similarity index), and a certain microorganism can be basically determined if the similarity index SI is more than 0.9. The system is a microorganism identification system which is relatively quick and has rich bacteria library, and has been widely applied (Wu Hamni, Xujian, Wang Haizhen, etc. SherlockMIS system is applied to the research of soil bacteria identification. soil bulletin, 2006, 43(4): 642-647).
In addition, 16S rDNA identification method was also used for verification and comparison with Sherlock Microbiological Identity System (MIS). Taking LM-LZ strain total DNA as a template, carrying out PCR amplification by using a universal primer of a 16S rDNA gene, recovering and sequencing an obtained amplified fragment to determine the size of the amplified fragment, and carrying out homology comparison on a sequencing result and a sequence in GenBank by using BLAST software.
1.3.4 Observation of morphological characteristics of bacterial strains and measurement of physiological and biochemical characteristics
Inoculating the strain to a culture medium containing 30 g.L-1In a solid basic culture medium of potassium nitrate, observing the morphological characteristics of the strains by an electron microscope after 48 hours; taking the logarithmic phase of the growth of the purified strain to perform gram and crystal violet simple capsule staining and the like; the physiological and biochemical characteristics are determined according to the handbook of identification of common bacterial systems (Dongxu pearl, Chuia Miao Yin).
2. Results
2.1 isolation and screening of the strains
Separating, purifying and screening to obtain 1 strain of bacteria capable of effectively treating nitrogen-containing wastewater, named as LM-LZ, and the others at 30 g.L-1The degradability of strains (such as LM-LZq, LM-LZs and the like) capable of growing on a solid basic culture medium of potassium nitrate is degraded to different degrees, the strains are cultured for 3 days, and the removal rates of total nitrogen, ammonia nitrogen, nitrate nitrogen and nitrite nitrogen of pollutants in nitrogen-containing wastewater by LM-LZ are respectively found to be96.71%, 97.42%, 95.93% and 96.78%.
2.2 identification of the strains
2.2.1 basic morphology and physiological and biochemical characteristics of LM-LZ strain
The bacillus brevis is a bacillus brevis, has rough surface, pilus and no cytopenia, has small capsule and gram-positive bacteria, has the size of (1.2-1.7) Mumx (0.4-0.7) Mum, and has circular bacterial colony, convex and smooth surface, complete edge, white color and opacity on a solid culture medium. The reaction of the formation test of the acetyl methyl methanol (v-p) is positive, the indole test is positive, the gelatin can be liquefied, and other physiological and biochemical characteristics are shown in the table 1.
TABLE 1 physiological and biochemical characteristics of WeideMannich Bacillus (Bacillus Wiedmannii) LM-LZ
Note: + positive reaction; negative reaction
2.2.2 identification of Strain LM-LZ
(1) Identification of LM-LZ by Sherlock MIS system
According to a fatty acid map generated by qualitative and quantitative analysis of the LM-LZ fatty acid by a Sherlock MIS software system, comparing with a Library database, preliminarily identifying that the LM-LZ is Wedney Mannich Bacillus (Bacillus Wiedmannii) and the similarity index SI (similarity index) is 0.993.
(2) Verification of LM-LZ by 16S rDNA identification method
DNA extraction results
PCR amplification was performed using the total DNA of the LM-LZ strain as a template and a universal primer for the 16S rDNA gene to obtain 1 fragment of about 1kb (FIG. 2). The amplified fragment is recovered and sequenced to confirm that the size is 1375bp (SEQ ID NO. 1).
II.16S rDNA Gene PCR amplification and sequence analysis
The homology comparison of the sequencing result and the sequence in GenBank by using BLAST software shows that the LM-LZ strain has high homology with Wedner Mannich Bacillus (Bacillus Wiedmannii), the homology is close to 100 percent, and the genetic distance is very close.
From the combination of the physiological and biochemical properties of LM-LZ, Sherlock MIS (MIDI identification system) and the evolutionary analysis of 16S rDNA system, it can be seen that the strain belongs to Wedner Mannich Bacillus (Bacillus Wiedmannii).
3 conclusion
1 high-efficiency strain LM-LZ (CGMCC No.18768) capable of effectively treating nitrogen-containing wastewater is obtained by separating biogas slurry oxidation pond sludge from water of a domestic pig farm in north town of Yu-watershed in Xinxi province, and is identified as Bacillus Wiedmannii (Bacillus Wiedmannii) through a Sherlock MIS system and 16S rDNA.
LM-LZ was deposited with the following information:
the preservation name is: wideher mannich Bacillus Wiedmannii, depository: china general microbiological culture Collection center, preservation Address: the microbial research institute of the national academy of sciences No. 3, Xilu No.1, Beijing, Chaoyang, Beijing, with the preservation number: CGMCC No.18768, preservation date: 2019, 10 and 30.
Example 2 detection of the Nitrogen-containing wastewater treatment Performance by the Strain
1 materials and methods
1.1 media and reagents
Basic culture medium: the same as in example 1.
Nitrogen-containing wastewater: from 20.48 g.L-1Sodium acetate, 1.44 g.L-1Potassium nitrate, 0.98 g.L-1Sodium nitrite, 0.94 g.L-1Ammonium sulfate, 0.5 g.L-1K2HPO4·3H2O and 0.03 g.L-1MgSO4·7H2O, pH 7.0. Wherein the total nitrogen is 593.0 mg.L-1194.8 mg. L of ammonia nitrogen-1202.2 mg.L of nitrate nitrogen-1196.0 mg.L of nitrite nitrogen-1。
1.2 Performance of the Strain on Nitrogen-containing wastewater treatment
A single purified Wedner mannich Bacillus (Bacillus Wiedmannii) LM-LZ strain is subjected to foundation treatmentThe culture medium propagation comprises the following specific steps: inoculating Bacillus WeideMannich (Bacillus Wiedmannii) LM-LZ into 100mL of basal medium, and inoculating at 30 deg.C for 180r min-1Culturing on a shaking table for 7 days; the obtained culture solution is transferred into 1000mL of basal culture medium according to the inoculation amount of 10 percent (volume percent), and the culture is continued for 3 days under the same culture conditions; obtaining bacterial liquid.
Measured as bacterial load OD415nmInoculating 0.2 to 100mL of nitrogen-containing wastewater, namely inoculating the bacterial liquid (about 10mL) of the amplified Wedner Mannich Bacillus (Bacillus Wiedmannii) LM-LZ into a 250mL triangular flask filled with 100mL of nitrogen-containing wastewater, wherein the bacterial amount OD in the wastewater415nm0.2; using nitrogen-containing waste water without inoculation as control, at 30 deg.C and 180 r.min-1The aerobic culture is carried out for 3 days on a constant temperature shaking table. After the culture is finished, the removal rate of the strain LM-LZ on total nitrogen, ammonia nitrogen, nitrate nitrogen and nitrite nitrogen in the nitrogen-containing wastewater is measured according to the method of example 1.
2. Results
The strain LM-LZ has pH of 7.0 and OD415nm0.2, rotation speed 180 r.min-1And under the aerobic environment, the removal rates of total nitrogen, ammonia nitrogen, nitrate nitrogen and nitrite nitrogen in the nitrogenous wastewater are respectively 96.71%, 97.42%, 95.93% and 96.78%. The total nitrogen, ammonia nitrogen, nitrate nitrogen and nitrite nitrogen residual concentrations are respectively 19.56 mg.L-1、5.02mg·L-1、8.23mg·L-1And 6.31 mg. L-1Can reach the first class B limit standard (total nitrogen concentration is 20 mg.L) of the pollutant discharge standard (GB18918-2002) of the urban sewage treatment plant-1The ammonia nitrogen concentration is 8 mg.L-1) And discharging shows that the separated and screened Wedner Mannich Bacillus (Bacillus Wiedmannii) is an efficient strain capable of effectively treating the nitrogen-containing wastewater, and has certain application potential for controlling the influence of the nitrogen-containing wastewater on the environment.
Comparative experiment:
the remaining strains obtained during the screening process of the present invention and other existing Bacillus species with a certain denitrification (as described in Table 2 below) were tested according to the method described in example 2 after replacing LM-LZ, and the results obtained are compared with those of LM-LZ as described in Table 2 below. The found Wedner Mannich Bacillus (Bacillus Wiedmannii) LM-LZ is an efficient strain capable of effectively treating nitrogen-containing wastewater, can efficiently remove total nitrogen, ammonia nitrogen, nitrate nitrogen and nitrite nitrogen in the wastewater at the same time, has certain application potential for controlling the influence of the nitrogen-containing wastewater on the environment, and other similar genera do not have the capacity or have weaker capacity.
TABLE 2
Example 3: detection of treatment performance of strain on nitrogen-containing wastewater in pig farm
1 materials and methods
1.1 media and reagents
Basic culture medium: the same as in example 1.
Nitrogen-containing wastewater produced by pig farms: pH6.8, total nitrogen 570.0 mg.L-1414.0 mg/L of ammonia nitrogen-1110.0 mg/L of nitrate nitrogen-146.0 mg.L of nitrite nitrogen-1。
The method is the wastewater leached in the pig farm water flushing fence breeding mode and the manure soaking mode.
1.2 treatment of Nitrogen-containing wastewater from pig farms with the Strain
The "nitrogen-containing wastewater" in example 2 was replaced with "nitrogen-containing wastewater from pig farm", and the rest was the same as example 2.
2. Results
The strain LM-LZ has pH of 6.8 and OD415nm0.2, rotation speed 180 r.min-1And under the aerobic environment, the removal rates of the total nitrogen, the ammonia nitrogen, the nitrate nitrogen and the nitrite nitrogen in the nitrogenous wastewater of the pig farm are respectively 98.33%, 98.73%, 97.26% and 97.29%. The residual concentrations of the total nitrogen, the ammonia nitrogen, the nitrate nitrogen and the nitrite nitrogen of the wastewater are respectively 9.52 mg.L-1、5.26mg·L-1、3.01mg·L-1And 1.25 mg. L-1Can reach the first class B limit standard (total nitrogen concentration is 20 mg.L) of the pollutant discharge standard (GB18918-2002) of the urban sewage treatment plant-1The ammonia nitrogen concentration is 8 mg.L-1) Discharge, which shows widemann separated and screened in the researchBacillus subtilis (Bacillus Wiedmanii) is a high-efficiency strain capable of effectively treating nitrogen-containing wastewater in a pig farm, and has certain application potential for controlling the influence of the nitrogen-containing wastewater in the pig farm on the environment.
Example 4 detection of the Performance of the strains on the treatment of Nitrogen-containing wastewater from synthetic pharmaceutical factories
1 materials and methods
1.1 media and reagents
Basic culture medium: the same as in example 1.
Nitrogen-containing wastewater generated by a synthetic pharmaceutical factory: pH7.0, total nitrogen 427.0 mg.L-1173.0 mg/L of ammonia nitrogen-1237.0 mg/L of nitrate nitrogen-1Nitrite nitrogen 17.0 mg.L-1。
1.2 treatment of Nitrogen-containing wastewater from synthetic pharmaceutical factory with the strain
The "nitrogen-containing wastewater" in example 2 was replaced with "nitrogen-containing wastewater from a synthetic drug factory", and the rest was the same as example 2.
2. Results
The strain LM-LZ has pH of 7.0 and OD415nm0.2, rotation speed 180 r.min-1And under the aerobic environment, the removal rates of total nitrogen, ammonia nitrogen, nitrate nitrogen and nitrite nitrogen in the nitrogen-containing wastewater of the synthetic pharmaceutical factory are respectively 96.49%, 96.83%, 96.27% and 96.3%. The residual concentrations of total nitrogen, ammonia nitrogen, nitrate nitrogen and nitrite nitrogen are respectively 14.95 mg.L-1、5.48mg·L-1、8.84mg·L-1And 0.63 mg. L-1Can reach the first class B limit standard (total nitrogen concentration is 20 mg.L) of the pollutant discharge standard (GB18918-2002) of the urban sewage treatment plant-1The ammonia nitrogen concentration is 8 mg.L-1) And (4) discharging, which shows that the separated and screened Wedner Mannich Bacillus (Bacillus Wiedmannii) is an efficient strain capable of effectively treating nitrogen-containing wastewater of a synthetic pharmaceutical factory, and has certain application potential for controlling the influence of the nitrogen-containing wastewater of the synthetic pharmaceutical factory on the environment.
Finally, it is also noted that the above-mentioned lists merely illustrate a few specific embodiments of the invention. It is obvious that the invention is not limited to the above embodiments, but that many variations are possible. All modifications which can be derived or suggested by a person skilled in the art from the disclosure of the present invention are to be considered within the scope of the invention.
Sequence listing
<110> New Yuhe-Biotech Co., Ltd
<120> Wedner bacillus LM-LZ separated from oxidation pond of pig farm and application thereof
<160>1
<170>SIPOSequenceListing 1.0
<210>1
<211>1375
<212>DNA
<213>2 Ambystoma laterale x Ambystoma jeffersonianum
<400>1
cccaccgact tcgggtgtta aaactctcgt ggtgtgacgg gcggtgtgta caaggcccgg 60
gaacgtattc accgcggcat gctgatccgc gattactagc gattccagct tcatgtaggc 120
gagttgcagc ctacaatccg aactgagaac ggttttatga gattagctcc acctcgcggt 180
cttgcagctc tttgtaccgt ccattgtagc acgtgtgtag cccaggtcat aaggggcatg 240
atgatttgac gtcatcccca ccttcctccg gtttgtcacc ggcagtcacc ttagagtgcc 300
caacttaatg atggcaacta agatcaaggg ttgcgctcgt tgcgggactt aacccaacat 360
ctcacgacac gagctgacga caaccatgca ccacctgtca ctctgctccc gaaggagaag 420
ccctatctct agggttttca gaggatgtca agacctggta aggttcttcg cgttgcttcg 480
aattaaacca catgctccac cgcttgtgcg ggcccccgtc aattcctttg agtttcagcc 540
ttgcggccgt actccccagg cggagtgctt aatgcgttaa cttcagcact aaagggcgga 600
aaccctctaa cacttagcac tcatcgttta cggcgtggac taccagggta tctaatcctg 660
tttgctcccc acgctttcgc gcctcagtgt cagttacaga ccagaaagtc gccttcgcca 720
ctggtgttcc tccatatctc tacgcatttc accgctacac atggaattcc actttcctct 780
tctgcactca agtctcccag tttccaatga ccctccacgg ttgagccgtg ggctttcaca 840
tcagacttaa gaaaccacct gcgcgcgctt tacgcccaat aattccggat aacgcttgcc 900
acctacgtat taccgcggct gctggcacgt agttagccgt ggctttctgg ttaggtaccg 960
tcaaggtgcc agcttattca actagcactt gttcttccct aacaacagag ttttacgacc 1020
cgaaagcctt catcactcac gcggcgttgc tccgtcagac tttcgtccat tgcggaagat 1080
tccctactgc tgcctcccgt aggagtctgg gccgtgtctc agtcccagtg tggccgatca 1140
ccctctcagg tcggctacgc atcgttgcct tggtgagccg ttacctcacc aactagctaa 1200
tgcgacgcgg gtccatccat aagtgacagc cgaagccgcc tttcaatttc gaaccatgca 1260
gttcaaaatg ttatccggta ttagccccgg tttcccggag ttatcccagt cttatgggca 1320
ggttacccac gtgttactca cccgtccgcc gctaacttca taagagcaag ctctt 1375
Claims (4)
1. Wedner mannich Bacillus (Bacillus Wiedmannii) LM-LZ separated from bottom mud of a pig farm oxidation pond is characterized in that: is Wedner Mannich Bacillus Bacillus Wiedmannii with a preservation number: CGMCC No. 18768.
2. The Wedner Mannich Bacillus (Bacillus Wiedimannii) LM-LZ according to claim 1, characterized in that: the 16S rDNA gene sequence of the strain is shown in SEQ ID NO. 1.
3. Use of Bacillus widmansonii (Bacillus Wiedmannii) LM-LZ according to claim 1 or 2, characterized in that: treating the nitrogen-containing wastewater.
4. Use of Brevibacillus brevis (Brachymonas sp) LM-R according to claim 3, characterized in that: belongs to heterotrophic nitrification-aerobic denitrifying bacteria, and can effectively remove the total nitrogen in the wastewater.
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