CN113215027B - Alcaligenes aquaticum AS1 and application thereof in sewage treatment - Google Patents

Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to the technical field of microbial wastewater treatment. The invention discloses an Alcaligenes aquaticus AS1 with the preservation number of CGMCC No. 21282. The strain is preserved in China general microbiological culture Collection center (CGMCC) at 12 months and 2 days in 2020, is called CGMCC for short, and has the address of No. 3 of Xilu No.1 of Beijing, Chaoyang, and the preservation number of CGMCC No. 21282. The strain can realize the synchronous removal of ammonia nitrogen, total nitrogen and COD under aerobic conditions.

Description

Alcaligenes aquaticum AS1 and application thereof in sewage treatment

Technical Field

The invention belongs to the technical field of microorganisms, and particularly relates to the technical field of microbial wastewater treatment.

Background

Main pollution index substances with overproof detection in fresh water resources (including surface water, underground water and the like of rivers, lakes, various large hydraulic projects and the like) in China comprise Chemical Oxygen Demand (COD) and ammonia Nitrogen (NH)₄-N), Total Nitrogen (TN), and Total Phosphorus (TP), among others. The nitrogen element has various existing pollutant forms and is high in removal difficulty, and when nitrogen-containing pollutants such as ammonia and nitrite with high concentration are discharged into the environment, on one hand, eutrophication of a water body can be caused, and on the other hand, biological toxicity can be generated to various organisms, so that the denitrification theory and the technology are always research hotspots in the field of sewage treatment.

Biological denitrification is the most economic and effective way for removing nitrogen in sewage at present. The traditional biological denitrification process comprises two processes of aerobic autotrophic nitrification and anaerobic heterotrophic denitrification, but has the following defects: (1) the autotrophic nitrifying bacteria have slow growth, poor environmental adaptability and weak impact load resistance, and are easily inhibited by high-concentration ammonia nitrogen and nitrite nitrogen. (2) The two reactions of nitrification and denitrification can not be unified in time and space, and the investment and operation cost is increased.

Heterotrophic nitrification and aerobic denitrification are a novel denitrification process. The device can realize simultaneous nitrification and denitrification in one reactor, complete synchronous removal of carbon and nitrogen pollutants, shorten the reaction period, save the space and reduce the operation cost. At present, a plurality of heterotrophic nitrification-aerobic denitrification strains are separated and screened, for exampleBacillus subtilis、 Pseudomonas stutzeri、Klebsiella pneumoniae、Alcaligenes faecalisAnd the like. However, most of the strains separated at present have harsh suitable growth conditions, limited application to denitrification of special types of wastewater (such as acid-base wastewater, high-salt wastewater and high-ammonia nitrogen wastewater), and high-efficiency Alcaligenes aquaticum (for example, Alcaligenes aquaticum) with salt resistance, acid-base resistance and high-concentration ammonia nitrogen removal and high removal rate for pig-raising wastewater COD and ammonia nitrogenAlcaligenes aquatilis) No report is found.

Disclosure of Invention

The invention aims to provide a strain of alcaligenes aquaticus (A. natans.)Alcaligenes aquatilis) AS1 with the preservation number of CGMCC number 21282. The strain is preserved in China general microbiological culture Collection center (CGMCC) at 12 months and 2 days in 2020, is called CGMCC for short, and has the address of No. 3 of West Lu No.1 of Beijing, Chaoyang, and the preservation number of CGMCC number 21282.

The invention has the beneficial effects that:

1. the invention provides an alcaligenes aquaticus (Alcaligenes aquatilis) AS1 has good effect in the aspect of microbial treatment of domestic sewage, livestock and poultry breeding sewage and other types of sewage, has good removal effect on ammonia nitrogen and total nitrogen, and can remove most of COD in water body, thereby realizing the purpose of removing most of COD in water bodyThe sewage is synchronously denitrified and decarbonized, so that the denitrification process flow is greatly simplified, and the treatment space and the cost are saved.

2. The strain AS1 has wide pH adaptability, and can effectively remove ammonia nitrogen within the pH range of 5-11.5.

3. The strain AS1 has salt tolerance, the ammonia nitrogen removal rate can reach more than 90% under the salt concentration of 40 g/L, and when the salt concentration is 60 g/L, the ammonia nitrogen removal rate still reaches 24%.

4. The strain AS1 provided by the invention can realize the rapid denitrification treatment of high-concentration ammonia nitrogen sewage, wherein the ammonia nitrogen removal rate reaches 96.2%, the TN removal rate reaches 76.3%, and the COD removal rate reaches 70% after aerobic treatment for 72 hours in the actual pig raising wastewater with high ammonia nitrogen concentration (1300 mg/L).

Drawings

FIG. 1 shows Alcaligenes aquaticum (A)Alcaligenes aquatilis) Growth and heterotrophic nitrification profiles of AS 1.

FIG. 2 shows Alcaligenes aquaticum (A)Alcaligenes aquatilis) Growth and denitrification profiles of AS1 at different pH.

FIG. 3 shows Alcaligenes aquaticum: (Alcaligenes aquatilis) Growth and denitrification profiles of AS1 at different salinity.

FIG. 4 shows Alcaligenes aquaticum: (Alcaligenes aquatilis) And the denitrification performance of AS1 in sterilized domestic sewage.

FIG. 5 shows Alcaligenes aquaticum (A)Alcaligenes aquatilis) Denitrification performance of AS1 in sterilized pig wastewater.

FIG. 6 shows Alcaligenes aquaticum (A)Alcaligenes aquatilis) AS1 denitrification performance diagram in the actual pig-raising wastewater with high ammonia nitrogen concentration.

FIG. 7 shows Alcaligenes aquaticum (A)Alcaligenes aquatilis) AS1 denitrification performance in high ammonia nitrogen concentration actual pig wastewater.

Detailed Description

The media used in the examples are as follows:

LB culture medium: 5 g of yeast extract, 10 g of tryptone, 10 g of sodium chloride and 1L of distilled water.

Heterotrophic nitrification culture medium: 4 g of sodium chloride, 2.66 g of disodium hydrogen phosphate, 1 g of monopotassium phosphate, 4.41 g of potassium citrate, 0.38 g of ammonium chloride, 3 mL of trace element solution and 1L of distilled water, and the pH value is natural.

Solution of trace elements: 3 g of magnesium sulfate heptahydrate, 3 g of manganese sulfate, 3 g of magnesium sulfate heptahydrate, 1.12 g of boric acid, 0.3 g of ferrous sulfate heptahydrate, 0.6 g of calcium chloride dihydrate and 1L of distilled water.

Example 1

And (3) separating and identifying strains:

enrichment culture: the sample is aerobic activated sludge collected from a certain sewage treatment plant in Beijing by Cao Shi ahead in 2020 and 4 months, 20 mL of fully mixed activated sludge is taken and placed in 180 mL of 0.2% sodium chloride solution for suspension, 5 mL of suspension is taken and placed in a 250 mL conical flask filled with 100 mL of heterotrophic nitrification culture medium, and domestication enrichment culture is carried out at 30 ℃ and 180 rpm. The acclimation takes 48 h as a period. After each period, 10 mL of the enrichment solution is added into a new 100 mL of heterotrophic nitrification culture medium to continue enrichment culture, and the enrichment culture is continuously carried out for 5 periods. During which the removal of ammonia nitrogen from the culture broth was examined.

Collecting culture solution after 5 generations enrichment, and adding sterile water according to the ratio of 10^-3-10^-7The method comprises the following steps of (1) diluting in different proportions in a gradient manner, taking 100 mu L of diluent from each gradient, coating the diluent on a heterotrophic nitrification solid culture medium, placing the culture medium in a biochemical incubator for 48 hours at 30 ℃, observing the growth condition of bacterial colonies, selecting single bacterial colonies with different forms from the culture medium, respectively scribing on the heterotrophic nitrification solid culture medium by using a plate scribing method, and culturing for 48 hours under the same condition. Then selecting single colony to perform multi-time partition streak purification until obtaining pure strain, placing the purified strain in 20% glycerol, and freezing and preserving at-80 ℃ in a refrigerator for later use.

Strain screening: respectively picking purified strains by using an inoculating loop, inoculating the strains into a sterilized LB liquid culture medium, culturing for 24 hours at 30 ℃ under the condition of 180 rpm, sucking a certain amount of bacterial liquid according to the inoculation amount of 5% (v/v), centrifuging for 5 minutes under the condition of 4000 rpm, collecting thalli, washing with sterile water, inoculating the thalli into a 150 mL conical flask filled with 50 mL of heterotrophic nitrification culture medium, and inoculating the thalli into a 18-mL conical flask at 30 ℃ under the condition of 18 rpmCulturing for 72 h in a 0 rpm shaker, and sampling for 12 h, 24 h, 36 h, 48 h, 60 h and 72 h to detect NH in the culture solution₄N, TN and COD content. Screening out bacterial strains with heterotrophic nitrification function.

The heterotrophic nitrifying bacteria AS1 are obtained through the separation and purification process, the bacteria are short-rod-shaped in morphological characteristics, the size of the bacteria is (0.4-0.6) Mumx (0.8-1.1) Mum, no spores are produced, the bacterial colonies are circular, the edges of the bacterial colonies are neat and smooth, the bacterial colonies are faint yellow on an LB culture medium, and the bacterial colonies are gram-negative aerobic bacteria.

The obtained strain is subjected to molecular biological identification, and sequencing comparison is carried out after bacterial 16S rDNA sequence (SEQ ID NO. 1) is subjected to PCR amplification.

The amplification primers were 27F: AGAGTTTGATCMTGGCTCAG, 1492R: TACGGYTACCTTGTTACGACTT, respectively;

the reaction system is as follows: 10 × Buffer 2 μ L, 2.5 mM dNTP 1.5 μ L, Primer 11 μ L, Primer 21 μ L, template 1 μ L, enzyme 0.3 μ L, water 13.2 μ L, and total volume 20 μ L;

the reaction conditions are as follows: pre-denaturation at 95 ℃ for 5 min, denaturation at 95 ℃ for 30 sec at 30 cycles, annealing at 55 ℃ for 30 sec, extension at 72 ℃ for 1.5 min, extension at 72 ℃ for 10 min, and heat preservation at 4 ℃ for forever.

And detecting the PCR product by agarose gel electrophoresis, and sequencing.

Sequencing in forward and reverse directions, splicing to obtain 16S rDNA sequence of the strain, performing Blast comparison on the sequence in NCBI database, and displaying the comparison result with Alcaligenes aquaticum (Alcaligenes aquaticum)Alcaligenes aquatilis) LMG 22996 has the highest homology of 99.57%, and the strain is named as Alcaligenes aquaticum (II)Alcaligenes aquatilis) AS1, hereinafter referred to herein AS strain AS 1.

Alcaligenes aquaticum (A), (B), (C)Alcaligenes aquatilis) AS1 has been deposited in China general microbiological culture Collection center (CGMCC) at 12.2.2020 for short, with the address of No. 3 Siro-Lu No.1 of Beijing Kogyo and the preservation number of CGMCC number 21282.

Example 2

Growth and heterotrophic nitrification characteristics of strain AS 1:

activating and culturing a strain AS1 in an LB liquid test tube for 24 h, absorbing a certain amount of bacterial liquid according to the inoculum size (v/v) of 2%, centrifuging for 5 min under the condition of 4000 rpm, collecting thalli, washing with sterile water, inoculating the thalli into a 150 mL conical flask filled with 50 mL heterotrophic nitrification culture medium (the initial ammonia nitrogen concentration is 100 mg/L, and C/N = 20), culturing for 72 h in a shaking table at 30 ℃ and 180 rpm, sampling every 4 h during the period, measuring the OD600 value, centrifuging the sample for 10 min at 10000 rpm, taking supernatant to detect COD and NH in a water sample, and taking the supernatant to detect the thalli₄-N、NO₂-N、NO₃-N, TN content.

The results are shown in the following table and in FIG. 1. Growth of the strain started at 4 h after inoculation and was substantially complete after 24 h, during which time NH was added₄The contents of-N, TN and COD gradually decreased. By 32 hours of treatment, the ammonia nitrogen removal rate reaches 98%, the COD removal rate is 90.5%, the TN removal rate is 70.8%, and finally a small amount of nitrite nitrogen (10.7 mg/L) and nitrate nitrogen (9.6 mg/L) are accumulated.

Example 3

Effect of pH on growth and denitrification of strain AS 1:

adjusting the pH of the heterotrophic nitrification culture medium to 3, 4, 5, 7, 8.5, 9.5, 10.5 and 11.5 respectively, activating the strain AS1 according to a conventional method, absorbing a certain amount of bacterial liquid according to the inoculum size (v/v) of 2 percent, centrifuging for 5 min at 4000 rpm, collecting thalli, washing with sterile water, inoculating into the heterotrophic nitrification culture medium after adjusting the pH, culturing for 72 h in a shaking table at 30 ℃ and 180 rpm, detecting the OD600 value under each pH condition finally, centrifuging the sample for 10 min at 10000 rpm, removing the thalli, taking the supernatant to detect COD and NH in a water sample₄-the content of N.

The results are shown in the following table and fig. 2. The strain AS1 can grow well in the pH range of 7-10.5 and has heterotrophic nitrification, ammonia nitrogen removal rate of 92-95% and COD removal rate of 87-91%. The strain AS1 has wide application prospect in sewage treatment with low or high pH.

pH	OD600	Ammonia nitrogen removal (%)	COD removal Rate (%)
				3	0.129	0.0	0.0
4	0.109	0.0	0.0
				5	0.131	0.0	4.0
7	0.784	95.0	91.0
				8.5	0.724	94.9	89.7
9.5	0.760	93.9	90.7
				10.5	0.749	92.8	87.6
11.5	0.043	21.5	5.0

Example 4

Effect of salinity on growth and denitrification of strain AS 1:

adjusting the salinity of the heterotrophic nitrification culture medium to 5, 20, 40, 60 and 80 g/L by using NaCl, activating a strain AS1 according to a conventional method, absorbing a certain amount of bacterial liquid according to the inoculum size (v/v) of 2 percent, centrifuging for 5 min at 4000 rpm, collecting thalli, washing with sterile water, inoculating the thalli into the heterotrophic nitrification culture medium after adjusting the salinity, culturing for 72 h in a shaking table at 30 ℃ and 180 rpm, detecting the OD600 value under each pH condition finally, centrifuging the sample for 10 min at 10000 rpm, removing the thalli, taking the supernatant, and detecting NH in a water sample₄-N content.

The results are shown in the following table and fig. 3. The strain AS1 can perform better growth and heterotrophic nitrification within the salinity range of 5-60 g/L, wherein the ammonia nitrogen removal rate is over 90% when the salinity is 5-40 g/L, and the ammonia nitrogen removal rate is still 24% when the salinity is 60 g/L, which indicates that the strain AS1 has wide application prospect in sewage treatment with higher salinity.

NaCl (g/L)	OD600	Ammonia nitrogen removal (%)
			5	0.770	97.8
20	0.974	98.8
			40	1.297	90.1
60	0.410	24.2
			80	0.044	2.0

Example 5

The denitrification performance of the strain AS1 in sterilized domestic sewage is AS follows:

the C/N ratio of actual domestic sewage (initial ammonia nitrogen content is about 100 mg/L) is adjusted to about 20 (initial COD content is about 1980 mg/L) by adding citrate, then the sterilization treatment is carried out, the strain AS1 is inoculated into the sterilized domestic sewage according to the proportion of 2% (v/v) after being activated conventionally, aerobic treatment is carried out in a shaking table at 30 ℃ and 180 rpm, and the ammonia nitrogen, TN and COD content in the sewage are detected every 24 h, and the results are shown in the following table and figure 4.

The result shows that the strain AS1 can remove most of ammonia nitrogen and all COD in the domestic sewage within 24 hours after inoculation, the total nitrogen removal rate reaches more than 70%, the ammonia nitrogen removal rate reaches 98.7% after 48 hours, and the total nitrogen removal rate reaches 77.5%, which indicates that the strain AS1 has good application prospect in domestic sewage denitrification treatment with low ammonia nitrogen concentration.

Example 6

The denitrification performance of the strain AS1 in the sterilized pig wastewater is AS follows:

the pretreatment method of the laboratory pig raising wastewater comprises the steps of uniformly mixing fresh pig urine collected from a certain pig farm of great Kyoto, Beijing with tap water according to the proportion of 1:4, adding fresh pig manure with the mass fraction of 5%, uniformly stirring, standing for 24 hours, and taking the upper liquid as the laboratory pig raising wastewater for later use.

The C/N ratio of the actual pig raising wastewater (the initial ammonia nitrogen content is about 465 mg/L) is adjusted to about 20 (the initial COD content is about 11700 mg/L) by adding citrate, the sterilization treatment is carried out, the strain AS1 is inoculated into the sterilized pig raising wastewater according to the proportion of 5% (v/v) after being activated conventionally, aerobic treatment is carried out in a shaker at 30 ℃ and 180 rpm, the ammonia nitrogen, TN and COD content in the sewage are detected every 36 h, and the results are shown in the following table and figure 5.

The results show that the ammonia nitrogen content is reduced from 465 mg/L to 118 mg/L, the COD is reduced from 11700 mg/L to 6613 mg/L, the ammonia nitrogen is reduced to 10.4 mg/L from 72 h, the COD is reduced to 3438 mg/L, the ammonia nitrogen removal rate reaches 97.8% in 72 h, the COD removal rate is 70.6% and the TN removal rate reaches 73% in 36 h after the inoculation of the strain AS 1. The strain AS1 has good application prospect in denitrification treatment of livestock and poultry breeding sewage with high ammonia nitrogen concentration.

Example 7

The denitrification performance of the strain AS1 in the actual pig raising wastewater with high ammonia nitrogen concentration is AS follows:

the C/N ratio of the actual pig raising wastewater (initial ammonia nitrogen content is about 420 mg/L) is adjusted to about 15 (initial COD content is 5800 mg/L) by adding citrate, the strain AS1 is inoculated into the pig raising wastewater according to the proportion of 5% (v/v) after being activated conventionally, meanwhile, a treatment group without the added strain AS1 is taken AS a comparison, aerobic treatment is carried out in a shaker at 30 ℃ and 180 rpm, and the ammonia nitrogen, TN and COD content in the sewage are detected every 12 hours, and the results are shown in the following table and figure 6.

According to the results, after the treatment for 12 hours, the ammonia nitrogen removal rate of the CK group is 17%, the TN removal rate is 19.2%, and the COD removal rate is 9.0%; while the removal rates of ammonia nitrogen, TN and COD of the treatment group 12 h inoculated with the strain AS1 are respectively 37.3%, 32.7% and 34.8%. And at 24 h, the removal rates of ammonia nitrogen, TN and COD of the CK group are respectively 53.2%, 45.8% and 71.8%, and the removal rates of ammonia nitrogen, TN and COD of the AS1 group are respectively 88.3%, 71.3% and 73.8%. When the treatment is carried out for 36 h, the ammonia nitrogen content of AS1 group is finally reduced to 23.9 mg/L, the removal rate is AS high AS 94.4%, and the removal rate is improved by 41.9% compared with that of CK group; the total nitrogen removal rate of the AS1 group is 70 percent, which is improved by about 18 percent compared with the CK group; and the final removal rate of COD has no obvious difference. In conclusion, the strain AS1 can remarkably improve the removal rate of ammonia nitrogen and TN in the pig-raising wastewater in a short time, and accelerate the degradation rate of COD.

Example 8

The denitrification performance of the strain AS1 in the actual pig raising wastewater with high ammonia nitrogen concentration is AS follows:

when the pig raising wastewater for laboratories is prepared by using pig manure, pig urine and tap water, the ammonia nitrogen concentration of the wastewater is adjusted to 1300 mg/L by reducing the addition amount of the tap water, the C/N ratio of the wastewater is adjusted to about 15 (the initial COD content is about 17280 mg/L) by adding citrate, the strain AS1 is inoculated into the pig raising wastewater according to the proportion of 5% (v/v) after being activated conventionally, meanwhile, a treatment group without the strain AS1 is set AS a control, aerobic treatment is carried out in a shaking table at 30 ℃ and 180 rpm, and the ammonia nitrogen, TN and COD content in the wastewater are detected every 12 hours, and the results are shown in the following table and figure 7.

Time (h)

COD-CK (mg/L)

COD-AS1 (mg/L)

Ammonia nitrogen-CK (mg/L)

Ammonia nitrogen-AS 1 (mg/L)

Total nitrogen-CK (mg/L)

Total Nitrogen-AS 1 (mg/L)

0

17280±169.71

17280±169.71

1300±0

1300±0

1365±11.31

1365±11.31

12

16388±186.68

15993±278.6

1121.75±13.79

1087.75±9.55

1131±22.63

1026±111.72

24

12861±140.01

13240.5±347.19

983±41.72

755.75±49.85

1027.5±45.96

601.5±132.23

36

9793.5±208.6

8464.5±1489.87

810.75±15.2

374.25±48.44

743.5±41.72

546.25±27.22

48

5611±267.29

5967±43.84

687±26.16

168±30.41

764.75±60.46

491.25±27.22

60

4186.5±44.55

5392.5±108.19

669.25±18.03

65.25±1.06

699.5±96.17

335.5±53.74

72

4283.5±116.67

5348±73.54

673.75±7.42

49.25±25.81

568.75±63.99

324.25±69.65

According to the results, after 24 hours of treatment, the removal rates of ammonia nitrogen, TN and COD of the CK group are respectively 24.4%, 24.7% and 25.6%, while the removal rates of ammonia nitrogen, TN and COD of the treatment group inoculated with the strain AS1 are respectively 41.9%, 56.1% and 23.4%. And at 48 h, the removal rates of ammonia nitrogen, TN and COD of the CK group are 47.2%, 44% and 67.5% respectively, and the removal rates of ammonia nitrogen, TN and COD of the AS1 group are 87.1%, 64% and 65.5% respectively. When the treatment is carried out for 72 h, the ammonia nitrogen content of AS1 group is finally reduced to 49.3 mg/L, the removal rate is AS high AS 96.2%, and the removal rate is improved by 48% compared with that of CK group; the total nitrogen removal rate of the AS1 group was 76.3%, which is about 18% higher than that of the CK group. In conclusion, the strain AS1 can tolerate high-concentration ammonia nitrogen in the actual pig raising wastewater, the removal rate of the ammonia nitrogen and TN in the pig raising wastewater is obviously improved in a short time, and the application prospect in denitrification treatment of the actual pig raising wastewater is wide.

The above description is only an exemplary embodiment of the present invention, and is not intended to limit the scope of the present invention. Any equivalent changes and modifications that can be made by one skilled in the art without departing from the spirit and principles of the invention should fall within the protection scope of the invention.

Sequence listing

<110> institute of microbiology of Chinese academy of sciences

<120> Alcaligenes aquaticum AS1 and application thereof in sewage treatment

<160> 1

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1406

<212> DNA

<213> Alcaligenes aquaticus (Alcaligenes aquatilis)

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gtagcgccct ccttacggtt aggctaccta cttctggtga aacccactcc catggtgtga 60

cgggcggtgt gtacaagacc cgggaacgta ttcaccgcga cattctgatc cgcgattact 120

agcgattccg acttcacgca gtcgagttgc agactgcgat ccggactacg atcgggtttc 180

tgagattggc tccccctcgc gggttggcga ccctctgtcc cgaccattgt atgacgtgtg 240

aagccctacc cataagggcc atgaggactt gacgtcatcc ccaccttcct ccggtttgtc 300

accggcagtc tcattagagt gctcttgcgt agcaactaat gacaagggtt gcgctcgttg 360

cgggacttaa cccaacatct cacgacacga gctgacgaca gccatgcagc acctgtgttc 420

cggttctctt gcgagcactc ccaaatctct tcgggattcc agacatgtca agggtaggta 480

aggtttttcg cgttgcatcg aattaatcca catcatccac cgcttgtgcg ggtccccgtc 540

aattcctttg agttttaatc ttgcgaccgt actccccagg cggtcaactt cacgcgttag 600

ctgcgctact aaggcctaac ggccccaaca gctagttgac atcgtttagg gcgtggacta 660

ccagggtatc taatcctgtt tgctccccac gctttcgtgt ctgagcgtca gtattatccc 720

agggggctgc cttcgccatc ggtattcctc cacatatcta cgcatttcac tgctacacgt 780

ggaattctac ccccctctga catactctag ctcggcagtt aaaaatgcag ttccaaggtt 840

gagccctggg atttcacatc tttctttccg aaccgcctac acacgcttta cgcccagtaa 900

ttccgattaa cgcttgcacc ctacgtatta ccgcggctgc tggcacgtag ttagccggtg 960

cttattctgc agataccgtc agcagtatct cgtattagga gatacctttt cttctctgcc 1020

aaaagtactt tacaacccga aggccttcat catacacgcg ggatggctgg atcagggttt 1080

cccccattgt ccaaaattcc ccactgctgc ctcccgtagg agtctgggcc gtgtctcagt 1140

cccagtgtgg ctggtcgtcc tctcaaacca gctacggatc gttgccttgg tgagccttta 1200

ccccaccaac tagctaatcc gatatcggcc gctccaatag tgagaggttc cgaagaatcc 1260

ccccctttcc cccatagggc gtatgcggta ttagccactc tttcgagtag ttatcccccg 1320

ctactgggca cgttccgata tattactcac ccgtccgcca ctcgccacca agagagcaag 1380

ctctctcgtg ctgccgttcg actgca 1406

Claims (6)

1. Alcaligenes aquaticum (A), (B), (C)Alcaligenes aquatilis) AS1 with the preservation number of CGMCC number 21282; the bacillus is preserved in the China general microbiological culture Collection center.

2. A microbial preparation comprising the Alcaligenes aquaticum (A) according to claim 1 as an active ingredientAlcaligenes aquatilis）AS1。

3. The Alcaligenes aquaticum (A) of claim 1Alcaligenes aquatilis) Application of AS1 in removing COD, ammonia nitrogen or/and total nitrogen in sewage.

4. Use according to claim 3, wherein the sewage is domestic sewage or swine wastewater.

5. Use according to claim 3, wherein the pH of the effluent is in the range of 7 to 10.5.

6. Use according to claim 3, wherein the salinity of the effluent is in the range of 0-60 g/L.

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