CN105441348A - New bacillus subtilis strain, microecological preparation and application - Google Patents

New bacillus subtilis strain, microecological preparation and application Download PDF

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Publication number
CN105441348A
CN105441348A CN201410460159.1A CN201410460159A CN105441348A CN 105441348 A CN105441348 A CN 105441348A CN 201410460159 A CN201410460159 A CN 201410460159A CN 105441348 A CN105441348 A CN 105441348A
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nitrogen
water
bacillus subtilis
nitrite
ammonia nitrogen
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CN201410460159.1A
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CN105441348B (en
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胡婷
马青山
贠锦军
何增国
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Fujian Dabei nonghuayou Aquatic Technology Group Co.,Ltd.
JIANGSU DABEINONG AQUATIC TECHNOLOGY Co.,Ltd.
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Fujian Dabeinong Fisheries Science & Technology Co Ltd
TIANJIN CHANGNONG TECHNOLOGY Co Ltd
Beijing Dabeinong Technology Group Co Ltd
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Abstract

The invention belongs to the technical field of aquatic-product microecological preparations, and concretely relates to a n new bacillus subtilis strain, a microecological preparation and application. The disclosed bacillus subtilis H8-1 possesses the preservation number of CGMCC No. 9356, is capable of reducing harmful substances such as ammonia nitrogen and nitrite-state nitrogen in water in a pollution-free residue-free way, improving culture quality and improving culture economic benefit, and possesses promotion value. The degradation rates of bacillus subtilis H8-1 on ammonia nitrogen and nitrite-state nitrogen both with the concentration of 2.5 mg/L are respectively 81.72% and 99.70%.

Description

One bacillus subtilis novel bacterial strain, probiotics and application
Technical field
The invention belongs to aquatic products probiotics technical field, be specifically related to subtilis new strains, probiotics and application.
Background technology
In recent years, high-density, intensive aquaculture progressively replace tradition and raise pattern scattered, greatly improve output and the economic benefit of aquaculture.But people, improving constantly cultivation density with while pursuing cultured output, also result in a series of common problem.Excessive bait throwing in, causes mixed bait utilization ratio to reduce, causes Organic substance in water to increase; Rate of fertilizer application strengthens, and causes nitrogen in water, phosphorus surges; The Organic pollutants aquaculture wateies such as a large amount of ight soil; Water transparency declines, water quality deterioration.Organic matter decomposition excessive in water body not only consumes the dissolved oxygen in water body, water body is in anoxic condition, discharge many objectionable impuritiess, as ammonia nitrogen, nitrite nitrogen and hydrogen sulfide etc., affect the diversity of organism in water directly or indirectly, the cultivation of high-quality aquatic products kind is subject to the restriction of water quality and food organism.Control the key issue that cultivating pool water ecological setting Yi Shi China pond aquaculture develops in a healthy way, the control of water quality of aquaculture pond and purify extremely urgent.
At present, Chinese scholars conducts in-depth research, and probiotics is pollution-free with it, the feature such as noresidue and multifunctionality, and producing bacillus subtilis product exist with endosporic form, very strong to the resistibility of the poor environments such as high temperature, low temperature, high salinity, there is ability and the inulinase-producing activity of degradation of ammonia nitrogen and nitrite nitrogen significantly.At present, genus bacillus is mainly present in the production phase as the technical problem of falling nitrogen bacterium, also has in product practical application effect.Although bacterial strain a lot, the effect difference of bacterial strain is large, actual effective little.
Summary of the invention
For solving the problem, the invention provides a bacillus subtilis novel bacterial strain and probiotics.
Subtilis in the present invention ( bacillussubtilis) H8-1, obtain for applicant is separated from the water of shrimp feed coefficient pond, Fujian, its biological property is as follows: bacterium colony smooth surface, translucent, and in dirty white, bacterium colony is circular, edge engrail; Colonial morphology is shaft-like, and size is 0.8 ~ 1.2 × 1.5 ~ 4.0um, and gemma form is oval to column, and be positioned at thalline central authorities or slightly inclined, after sporulation, thalline does not expand.Subtilis in the present invention and existing subtilis be different from can under 0 ‰, 3 ‰, 15 ‰ salt concentration conditions equal well-grown, 10-15 DEG C of low temperature can be tolerated, well-grown.
Subtilis in the present invention ( bacillussubtilis) H8-1, its gelatin reacting positive, freezing cow's milk, reduction nitrate, edwardsiella hoshinae, H 2s, V-P reacting positive, starch experiment is positive, and glucose fermentation produces acid not aerogenesis, aerobic, is suitable for 25 ~ 37 DEG C of growths.
Subtilis in the present invention ( bacillussubtilis) H8-1, its 16SrRNA nucleotide sequence is as shown in SEQIDNo.1.Put into Genbank to compare, result display with bacillussubtilisthe similarity of 16SrRNA sequence reach 99%.In conjunction with ne ar feature, physio-biochemical characteristics, growth conditions, determine genus bacillus ( bacillussp.) H8-1 is subtilis.
Subtilis in the present invention ( bacillussubtilis) H8-1, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on June 18th, 2014, be called for short CGMCC, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, deposit number is CGMCCNo.9356.
The present invention also provides the probiotics containing described bacterial strain.
Bacterial strain of the present invention can be prepared into fodder additives, also can add in conventional fodder additives, and therefore, the present invention also provides a kind of fodder additives containing described bacterial strain.
Described fodder additives of the present invention can add conventional Preblend to, and as in vitamin-mineral Preblend, therefore, the present invention also provides a kind of Preblend containing described fodder additives.
The present invention also provides described bacterial strain or the application of described probiotics in improvement aquaculture water.
Particularly, bacterial strain of the present invention or described probiotics are for the ammonia nitrogen of water body of degrading and/or nitrite nitrogen level.
Subtilis in the present invention ( bacillussubtilis) H8-1 is that aquaculture water improvement probiotics provides provenance, reduce objectionable impurities in water in pollution-free and mode that is noresidue and, as ammonia nitrogen and nitrite nitrogen, improve cultivation quality, improve fanning economics, there is promotional value.Subtilis H8-1 in the present invention to concentration be the ammonia nitrogen of 2.5mg/L, nitrite nitrogen degradation rate is respectively 81.72%, 99.70%.
Embodiment
Following examples for illustration of the present invention, but are not used for limiting the scope of the invention.
Embodiment 1
Get and come from Wuqing, Tianjin, Fujian, the water sample on fish pond, Jiangsu and the shrimp pool and 124, mud sample, get 1mL water sample or mud sample 1g in the 9mLLB liquid nutrient medium of 18ml test tube, 85 DEG C of water bath processing 10min.
Test tube after pyroprocessing being placed on temperature is 30 DEG C, obtains enrichment culture liquid under the aerobic condition of 180rpm after constant-temperature shaking culture 16h.
Adopt coubling dilution that enrichment culture liquid is carried out gradient dilution, coat on LB solid medium, be positioned in 30 DEG C of thermostat containers and carry out cultivation 20h, the difference list bacterium colony that picking flat board grows is rule, and purifying twice, obtain single bacterium colony of living, obtain 275 strain bacterium altogether.
By the single bacterium colony after purifying twice, with toothpick dibbling on fresh haemolysis plate, be positioned in 30 DEG C of thermostat containers and observe 24h, 48h, 72h, whether there is haemolysis circle, Capability Categories according to making hemocyte agar haemolysis: 1) α haemolysis: grass green haemolysis, there is the grass green colour circle of 1 ~ 2mm in periphery of bacterial colonies substratum, caused by methemoglobin; 2) β haemolysis: complete hemolysis, periphery of bacterial colonies forms a complete Clear & Transparent zone of hemolysis, is that bacteriogenic hemolysin makes caused by red corpuscle dissolves completely; 3) γ haemolysis: not haemolysis.Haemolysis circle bacterial strain will be had to eliminate.
To rule on Selective solid culture medium without hemolytic strain, well-grown bacterial strain LB liquid culture, at 30 DEG C of constant temperature enlarged culturing 16h.
Solid selective medium is two kinds: with NH 4 +-N be only nitrogen source ammonia nitrogen selective medium and with NO 2 --N is the nitrite nitrogen substratum of only nitrogen source;
Ammonia nitrogen Selective solid culture medium is that every L is by (NH 4) 2sO 40.5g, sodium succinate 2.17g, Vickers salts solution 50mL, the water composition of agar 20g and surplus, pH7.0-7.2,115 DEG C of autoclaving 20min, described Vickers salts solution is that every L is by K 2hPO 45.0g, MgSO 47H 2o2.5g, NaCl2.5g, FeSO 47H 2o0.05g, MnSO 44H 2the water composition of O0.05g and surplus;
Nitrite nitrogen Selective solid culture medium is that every L is by NaNO 20.5g, sodium succinate 2.17g, Vickers salts solution 50mL, the water composition of agar 20g and surplus, pH7.0-7.2,115 DEG C of autoclaving 20min, described Vickers salts solution is that every L is by K 2hPO 45.0g, MgSO 47H 2o2.5g, NaCl2.5g, FeSO 47H 2o0.05g, MnSO 44H 2the water composition of O0.05g and surplus.
Final acquisition well-grown 63 bacillus without haemolysis circle and on ammonia nitrogen and nitrite nitrogen Selective solid culture medium.
Nitrogen function falls in embodiment 2 high throughput assay bacterial strain, screening high efficient strain
In super clean bench, access in corresponding screening culture medium by 63 strain test strains bacterium liquid, inoculation final concentration is 5 × 10 5cFU/mL, repeat number is 3; Put into shaking table and carry out cultivation 30 DEG C, 180r/min, cultivate 48h; Uniform sampling is in 1.5mL centrifuge tube, and the centrifugal 3min of 12000rpm, gets supernatant 200 μ L and add 96 orifice plates;
When measuring nitrite nitrogen, add in 96 orifice plates after test sample supernatant liquor 200 μ L, every hole successively adds each 20 μ L of griess reagent A, B, in 550nm colorimetric estimation; When measuring ammonium nitrogen, every hole adds nessler reagent 20 μ L, in 450nm colorimetric estimation;
Screening culture medium is two kinds: ammonia nitrogen screening culture medium and nitrite nitrogen screening culture medium;
Ammonia nitrogen screening culture medium is that every 50mL is by (NH 4) 2sO 4mother liquor 50 μ l, the water composition of commercially available pulverizing shrimp material 0.05g and surplus, ammonium nitrogen concentration is: 2.5mg/l;
Nitrite nitrogen screening culture medium is that every 50mL is by NaNO 2mother liquor 50 μ l, the water composition of commercially available pulverizing shrimp material 0.05g and surplus, nitrite nitrogen concentration is: 2.5mg/l.
Ammonium sulfate liquor is for taking 9g (NH 4) 2sO 4add 1L and simulate shrimp pool water, now ammonium nitrogen concentration is about: 2.5g/l;
Sodium Nitrite mother liquor is for taking 3.75gNaNO 2add 1L and simulate shrimp pool water, now nitrite nitrogen concentration is about: 2.5g/l.
Choose ammonia nitrogen degradation rate higher than 30% 10 strain bacterium (being numbered A1-A10) or cultured water degradation rate higher than the 10 strain bacterium (being numbered Y1-Y10) of 70%, not inoculate any bacterial strain for negative control (CK), carry out replication.
Measuring method: in super clean bench, accesses in corresponding screening culture medium by test strains bacterium liquid, and inoculation final concentration is 5 × 10 5cFU/mL, repeat number is 3; Put into shaking table and carry out cultivation 30 DEG C, 180r/min, cultivate 24h, 48h; Uniform sampling is in 1.5mL centrifuge tube, and the centrifugal 3min of 12000r/min, gets supernatant 200 μ L and add 96 orifice plates.
When measuring nitrite nitrogen, add in 96 orifice plates after test sample supernatant liquor 200 μ L, every hole successively adds each 20 μ L of griess reagent A, B, in 550nm colorimetric estimation; During mensuration ammonia nitrogen, every hole adds nessler reagent 20 μ L, in 450nm colorimetric estimation.
Qualitative test: when measuring nitrite nitrogen, solution becomes pink, rose, orange or brown etc. indicates nitrate reductase, and react for the positive, namely more the bright degradation effect of elementary introduction is better for color; When measuring ammonium nitrogen, it is positive reaction that solution becomes orange, yellow etc., and shallow is good.
Quantitative assay: formulate nitrite nitrogen concentration (Y 1) and measure OD value (X 1) between reference standard curve and ammonia nitrogen concentration (Y 2) and measure OD value (X 2) between reference standard curve.
Screening culture medium comprises ammonia nitrogen screening culture medium and nitrite nitrogen screening culture medium;
Ammonia nitrogen screening culture medium: (NH 4) 2sO 4mother liquor 50 μ l, shrimp material 0.05g, distilled water 50mL, ammonia nitrogen concentration is about: 2.5mg/L, 121 DEG C of sterilizings.Ammonium sulfate liquor: take 9g (NH 4) 2sO 4add 1L distilled water, now ammonium nitrogen concentration is about: 2.5g/l.
Nitrite nitrogen screening culture medium: NaNO 2mother liquor 50 μ l, shrimp material 0.05g, distilled water 50mL, nitrite nitrogen concentration is about: 2.5mg/l, 121 DEG C of sterilizings.Sodium Nitrite mother liquor: take 3.75gNaNO 2add 1L distilled water, now nitrite nitrogen concentration is about: 2.5g/l.
Ge Lisishi reagent (Griess) A: Sulphanilic Acid 0.5g, dilute acetic acid (about 10%) 150mL, brown bottle lucifuge, 4 DEG C of preservations.Ge Lisishi reagent B: αnaphthylamine 0.1g, distilled water 20mL, dilute acetic acid (about 10%) 150mL, brown bottle lucifuge 4 DEG C preservation.
Nessler reagent: cash purchase, 4 DEG C of preservations.
As shown in Table 1 and Table 2, the average ammonia nitrogen degradation rate of H8-1 is 81.72% to result, and average nitrous degradation rate is 99.70%, and it is the highest to fall nitrogen ability.
Table 110 strain preferred strain ammonia nitrogen degradation rate comparative measurement (No. A10 is H8-1)
Bacterial strain number A1 A2 A3 A4 A5 A6 A7 A8 A9 A10 ck
24h ammonia nitrogen 34.096 30.000 19.789 45.702 37.728 39.985 59.574 21.277 75.487 79.572 0.011
48h ammonia nitrogen 45.897 35.786 38.931 56.971 40.302 44.818 31.405 40.626 61.079 83.873 0.441
Table 210 strain preferred strain nitrite nitrogen degradation rate comparative measurement (No. Y10 is H8-1)
Bacterial strain number Y1 Y2 Y3 Y4 Y5 Y6 Y7 Y8 Y9 Y10 ck
24h nitrous 83.121 74.374 90.671 92.332 75.629 74.506 90.467 95.714 92.196 99.603 0.002
48h nitrous 83.018 74.508 90.128 93.671 75.457 70.165 92.467 96.541 94.467 99.806 2.443
The subtilis H8-1 screened is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on June 18th, 2014, be called for short CGMCC, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, deposit number is CGMCCNo.9356.
Under the different salt concentration conditions of embodiment 3, H8-1 growing state compares
Use salinity to be respectively the water preparation LB solid medium of 0 ‰, 3 ‰, 15 ‰, with the toothpick dibbling H8-1 after sterilizing on the flat board of different salinity, be positioned in 30 DEG C of incubators and cultivate 16h, observe dull and stereotyped upper colony growth situation.Find after cultivating, the colony growth of dibbling on the flat board that three kinds of salinity are different is all good, and the bacterium colony size after grow comparatively unanimously, illustrates that H8-1 salt resistant character is strong.
LB substratum: Tryptones: 10g/L; Yeast extract 5g/L; Sodium-chlor 10g/L; Agar powder 15g/L.Be respectively the water of 0 ‰, 3 ‰, 15 ‰ by salometer preparation salinity, carry out the preparation of substratum.
Under the different salt concentration conditions of embodiment 4, nitrogen performance measurement falls in H8-1
The results are shown in Table 3, it is as shown in the table, and under 0 ‰, 3 ‰, 15 ‰ salinity, nitrogen performance difference to some extent falls in H8-1, but very little by the impact of salinity.
What table 3 measured H8-1 under different salinity falls nitrogen performance
Ammonia nitrogen screening culture medium: (NH 4) 2sO 4mother liquor 50 μ l, shrimp material 0.05mg, use the water (water of 0 ‰ salinity: distilled water of different salinity respectively; The water of 3 ‰ and 15 ‰ salinity: add sea salt in distilled water, measure with salometer, reach salinity and be respectively 3 ‰ and 15 ‰) 50mL, ammonia nitrogen concentration is about: 2.5mg/L, 121 DEG C of sterilizings.Ammonium sulfate liquor: take 9g (NH 4) 2sO 4add 1L distilled water, now ammonium nitrogen concentration is about: 2.5g/l.
Nitrite nitrogen screening culture medium: NaNO 2mother liquor 50 μ l, shrimp material 0.05mg, the water 50mL of different salinity, nitrite nitrogen concentration is about: 2.5mg/l, 121 DEG C of sterilizings.Sodium Nitrite mother liquor: take 3.75gNaNO 2add 1L distilled water, now nitrite nitrogen concentration is about: 2.5g/l.
Measuring method is with embodiment 2.
Embodiment 5 is H8-1 growing state under cryogenic
With the mono-bacterium colony of toothpick dibbling H8-1 after sterilizing on LB solid plate, being placed on temperature is respectively 24h in 10 DEG C, 15 DEG C, 30 DEG C incubators, observes dull and stereotyped upper colony growth situation.Find after cultivating, on the flat board grown in 10 DEG C with 15 DEG C of incubators, bacterium colony size is similar, be growth on 30 DEG C of incubator middle plateforms bacterium colony size 1/2, this illustrates that H8-1 also can normal growth and play and fall nitrogen function in the breeding environment that temperature is lower.
LB solid medium: Tryptones: 10g/L; Yeast extract 5g/L; Sodium-chlor 10g/L; Agar powder 15g/L.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the prerequisite not departing from the technology of the present invention principle; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Sequence table
<110> BeiJing DaBei farm Science Group Co., Ltd
Fujian great Bei agriculture Aqua Sciences Inc.
The prosperous agriculture science and technology limited Company in Tianjin
<120> mono-bacillus subtilis novel bacterial strain, probiotics and application
<130>
<160>1
<170>PatentInversion3.3
<210>1
<211>1424
<212>DNA
<213> subtilis (Bacillussubtilis)
<400>1
cttggcggctgctctaaaggttacctcaccgacttcgggtgttacaaactctcgtggtgt60
gacgggcggtgtgtacaaggcccgggaacgtattcaccgcggcatgctgatccgcgatta120
ctagcgattccagcttcacgcagtcgagttgcagactgcgatccgaactgagaacagatt180
tgtgggattggcttaacctcgcggtttcgctgccctttgttctgtccattgtagcacgtg240
tgtagcccaggtcataaggggcatgatgatttgacgtcatccccaccttcctccggtttg300
tcaccggcagtcaccttagagtgcccaactgaatgctggcaactaagatcaagggttgcg360
ctcgttgcgggacttaacccaacatctcacgacacgagctgacgacaaccatgcaccacc420
tgtcactctgcccccgaaggggacgtcctatctctaggattgtcagaggatgtcaagacc480
tggtaaggttcttcgcgttgcttcgaattaaaccacatgctccaccgcttgtgcgggccc540
ccgtcaattcctttgagtttcagtcttgcgaccgtactccccaggcggagtgcttaatgc600
gttagctgcagcactaaggggcggaaaccccctaacacttagcactcatcgtttacggcg660
tggactaccagggtatctaatcctgttcgctccccacgctttcgctcctcagcgtcagtt720
acagaccagagagtcgccttcgccactggtgttcctccacatctctacgcatttcaccgc780
tacacgtggaattccactctcctcttctgcactcaagttccccagtttccaatgaccctc840
cccggttgagccgggggctttcacatcagacttaagaaaccgcctgcgagccctttacgc900
ccaataattccggacaacgcttgccacctacgtattaccgcggctgctggcacgtagtta960
gccgtggctttctggttaggtaccgtcaaggtaccgccctattcgaacggtacttgttct1020
tccctaacaacagagctttacgatccgaaaaccttcatcactcacgcggcgttgctccgt1080
cagactttcgtccattgcggaagattccctactgctgcctcccgtaggagtctgggccgt1140
gtctcagtcccagtgtggccgatcaccctctcaggtcggctacgcatcgttgccttggtg1200
agccgttacctcaccaactagctaatgcgccgcgggtccatctgtaagtggtagccgaag1260
ccaccttttatgtttgaaccatgcggttcaaacaaccatccggtattagccccggtttcc1320
cggagttatcccagtcttacaggcaggttacccacgtgttactcacccgtccgccgctaa1380
catcagggagcaagctcccatctgtccgctcgactgcatgatac1424

Claims (6)

1. a bacillus subtilis ( bacillussubtilis) new strains, its preserving number is CGMCCNo.9356.
2. the probiotics containing bacterial strain described in claim 1.
3. the fodder additives containing bacterial strain described in claim 1.
4. the Preblend containing fodder additives described in claim 3.
5. the application of bacterial strain according to claim 1 in improvement aquaculture water.
6. apply as claimed in claim 5, it is characterized in that, for ammonia nitrogen and/or the nitrite nitrogen level of water body of degrading.
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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105969708A (en) * 2016-08-01 2016-09-28 浙江大学 Bacillus subtilis SC228 with nitration function and application thereof
CN108865940A (en) * 2018-07-17 2018-11-23 中国海洋大学 One plant of heterotrophic nitrification-aerobic denitrification bacillus and its composite bacteria preparation
CN112011486A (en) * 2020-09-09 2020-12-01 天津市农业科学院 Synchronous nitrification and denitrification bacillus subtilis K8 and application thereof
CN112553112A (en) * 2020-12-19 2021-03-26 武汉水之国环保科技有限公司 Denitrifying bacillus subtilis and application thereof in strain detoxification
CN113249273A (en) * 2021-06-30 2021-08-13 泸州老窖股份有限公司 Salt-tolerant bacillus subtilis and application thereof in high-salt ammonia nitrogen wastewater treatment

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CN103773722A (en) * 2014-01-16 2014-05-07 中国药科大学 Salt-tolerance bacillus subtilis with low-temperature biological deamination function and application of bacillus subtilis

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CN1603248A (en) * 2003-02-14 2005-04-06 中国科学院南京土壤研究所 Biological denitrification method for ammonium-nitrogen containing waste water and microbes thereof
CN103773722A (en) * 2014-01-16 2014-05-07 中国药科大学 Salt-tolerance bacillus subtilis with low-temperature biological deamination function and application of bacillus subtilis

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Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105969708A (en) * 2016-08-01 2016-09-28 浙江大学 Bacillus subtilis SC228 with nitration function and application thereof
CN105969708B (en) * 2016-08-01 2019-09-03 浙江大学 Has the bacillus subtilis SC228 and application thereof of nitrification function
CN108865940A (en) * 2018-07-17 2018-11-23 中国海洋大学 One plant of heterotrophic nitrification-aerobic denitrification bacillus and its composite bacteria preparation
CN108865940B (en) * 2018-07-17 2021-10-08 中国海洋大学 Heterotrophic nitrification-aerobic denitrification bacillus and composite bacterial preparation thereof
CN112011486A (en) * 2020-09-09 2020-12-01 天津市农业科学院 Synchronous nitrification and denitrification bacillus subtilis K8 and application thereof
CN112011486B (en) * 2020-09-09 2022-03-01 天津市农业科学院 Synchronous nitrification and denitrification bacillus subtilis K8 and application thereof
CN112553112A (en) * 2020-12-19 2021-03-26 武汉水之国环保科技有限公司 Denitrifying bacillus subtilis and application thereof in strain detoxification
CN113249273A (en) * 2021-06-30 2021-08-13 泸州老窖股份有限公司 Salt-tolerant bacillus subtilis and application thereof in high-salt ammonia nitrogen wastewater treatment
CN113249273B (en) * 2021-06-30 2022-03-08 泸州老窖股份有限公司 Salt-tolerant bacillus subtilis and application thereof in high-salt ammonia nitrogen wastewater treatment

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