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Acid-resistant alkali-resistant rapid-denitrification aerobic denitrifying bacterium, microbial inoculum and application of aerobic denitrifying bacterium and microbial inoculum

A technology of aerobic denitrification, acid and alkali resistance, applied in the field of microbial denitrification, can solve the problem of lack of biosafety test and drug resistance test of microbial strains, difficult high acid and alkaline industries, breeding wastewater, beneficial mechanism research Few problems, such as good water body organic carbon removal capacity, simplified wastewater treatment process, high aquatic biological safety effect

Active Publication Date: 2022-04-22
GUANGZHOU UNIVERSITY
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Problems solved by technology

[0005] In terms of acid and alkali resistance, Maoxia Chen et al. reported on May 4, 2014 a strain of Aeromonas isolated from the active sludge of the CASS reactor Bacteria (Aeromonas sp.) HN-02 (Maoxia Chen et al., Bioresource Technology.167:456-461 (2014)), this bacterial strain can play a role in reducing nitrogen at pH=2.3~10, at pH=2.3, the concentration of ammonia nitrogen is At 200mg / L, it still has a removal rate of 39.0% for 48 hours, and has strong resistance to strong acid and weak alkali. Its removal ability of ammonia nitrogen under high alkali conditions (pH>11) is inhibited, less than 30%; Chinese patent CN106754534A A strain of Pseudomonas aeruginosa (Pseudomonas aeruginosa) is disclosed, which still has denitrification ability under the condition of pH=4.5, and the nitrate removal rate reaches 90% (initial concentration 160mg / L) within 72h; In the case of ~7.71, nitrate nitrogen with an initial concentration of 150mg / L can be completely removed in 32 hours, but the removal rate of nitrate nitrogen for this strain is relatively slow, and it is cultured under optimal conditions, and it is a lagging period from 0 to 24 hours , enter logarithmic growth phase after 24h
However, there is little research on the beneficial mechanism of aerobic denitrifying bacteria in aquatic systems. At present, in the process of strain research and transition to practical application, biosafety tests and drug resistance tests on microbial strains are often lacking. , which may ignore the potential harmful effects of some microbial agents, so people still need to isolate and purify more efficient, stable and safe aerobic denitrifying strains for further biosafety and ecological risk assessment
[0007] To sum up, the existing technology still lacks high-efficiency nitrogen reduction capabilities, high biological safety, and the ability to simultaneously treat high-acid and high-alkaline nitrogen-containing wastewater. Aerobic denitrifying strains, it is difficult to use biological methods to treat high acidity and alkalinity industrial and aquaculture wastewater

Method used

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  • Acid-resistant alkali-resistant rapid-denitrification aerobic denitrifying bacterium, microbial inoculum and application of aerobic denitrifying bacterium and microbial inoculum
  • Acid-resistant alkali-resistant rapid-denitrification aerobic denitrifying bacterium, microbial inoculum and application of aerobic denitrifying bacterium and microbial inoculum
  • Acid-resistant alkali-resistant rapid-denitrification aerobic denitrifying bacterium, microbial inoculum and application of aerobic denitrifying bacterium and microbial inoculum

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Experimental program
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Effect test

Embodiment 1

[0045] Isolation and identification of the ZY-3 strain of the present invention

[0046] 1. Sample collection

[0047] Pseudomonas mutans ZY-3 of the present invention is cultivated in tilapia in Nanhai Tongwei Aquatic Technology Co., Ltd. (N: 22°50′34″, E: 113°57′25″) in Nanhai District, Foshan City, Guangdong Province The water samples and mud samples of the pond were screened and separated; the samples were collected according to the "mixed sample collection method" in the "Technical Specifications for Soil Environment Monitoring" (HJ / T 166-2004), and the plum blossom point sampling method was used to collect samples from the breeding ponds. Collect surface, middle and deep water and sediment in sterile sampling bags, refrigerate, transport and store at 4°C for later use;

[0048] 2. Preparation of medium and solution

[0049] (1) Trace element solution (g / L): EDTA50g, ZnSO 4 ·7H 2 O 5.02g, CuSO 4 ·5H 2 O 1.57g, FeSO 4 ·7H 2 O 5.0g, CoCl 2 ·6H 2 O 1.61g, (NH 4 ) ...

Embodiment 2

[0072] Specific application and environmental safety evaluation of ZY-3 bacterial strain and bacterial agent

[0073] (1) Fish toxicity test: select healthy zebrafish (Danio rerio) with a body length within the range of 3±1cm, and keep them in a large water body with continuous aeration for 30 days. During this period, they are fed normally and the water is changed regularly; the state is stable Afterwards, they were randomly assigned to 15L glass jars, and the experimental group (P.plecoglossicida ZY-3) adding the bacterial agent (bacteria liquid) of the ZY-3 bacterial strain and the control group (CRT) adding equal volume of sterile water were set, and each experiment Group 30 zebrafish, and set 3 replicates in each group; take the overnight cultured bacterial solution, centrifuge at 4000r / min for 5min, discard the supernatant, resuspend with sterile PBS buffer, repeat 1 or 2 times, and rinse with sterilized water Suspend, get OD according to the determined standard curve 6...

Embodiment 3

[0087] The growth of strain ZY-3 and bacterial agents under various high-concentration single nitrogen sources and the performance test for nitrogen removal

[0088] The high-concentration single nitrogen source medium (DM) was used as the basal medium, and a single inorganic nitrogen source NH was added respectively. 4 Cl(DMⅠ), NaNO 3 (DMⅡ), NaNO 2(DMⅢ) To test the nitrogen-reducing ability of Pseudomonas mutans ZY-3 under high-concentration nitrogen source, the amount added per liter of medium was 0.6036g, 0.9590g, 0.7790g respectively, and the culture conditions were 28°C, 180r / min , pH=7.0; the ZY-3 strain was inoculated in nutrient broth medium, cultivated at 30°C, 180r / min for 24h to the logarithmic phase, and then inoculated with 1% of the inoculum, respectively, into the above-mentioned reactions with different organic carbon sources In the nitrification medium, take the culture solution at 0h, 4h, 12h, 24h, 36h, and 48h to measure its OD 600 , 4000r / min, 5min low-s...

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Abstract

The invention belongs to the technical field of microbial denitrification, and discloses an acid-resistant and alkali-resistant aerobic denitrifying bacterium capable of rapidly denitrifying, a microbial agent and application thereof, the aerobic denitrifying bacterium is a Pseudomonas plecoglossa strain ZY-3, is preserved in Guangdong Microbiological Culture Collection Center (GDMCC) on January 4, 2022, and has a preservation number of GDMCC No: 62181; the strain and the inoculant have the heterotrophic nitrification and aerobic denitrification capabilities at the same time, the growth speed is high, the nitrogen reduction rate is high, and the strain and the inoculant can be respectively utilized and used as a unique inorganic nitrogen source for aerobic nitrification and denitrification nitrogen removal. The strain and the microbial inoculum can play a nitrification-denitrification role in a water body environment with the pH value of 5.5-10.5, and meanwhile, the strain and the microbial inoculum have good biological safety.

Description

technical field [0001] The invention relates to the technical field of microbial denitrification, in particular to an aerobic denitrification bacterium, in particular to an acid-resistant and alkali-resistant rapid denitrification aerobic denitrification bacterium, a bacterial agent and its application in biological denitrification treatment. Background technique [0002] China is a big country in the aquaculture industry. At present, my country's aquaculture model is high-density and intensive. This model is easy to operate, high in output, and good in benefits. However, it is easy to cause the accumulation of harmful substances such as ammonia nitrogen and nitrite in the aquaculture water, which is harmful to the environment. , humans and farmed animals all have certain hazards, which is not conducive to long-term development. At present, the effective denitrification methods for treating nitrogen-containing aquaculture tail water mainly include physical treatment, chemical...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C12R1/38C02F103/20C02F103/34C02F101/38C02F101/16
CPCC12N1/20C02F3/34C02F2103/20C02F2103/34C02F2101/38C02F2101/16C02F2209/02C02F2209/06Y02W10/10
Inventor 舒琥林浩澎马永浩孙慧明陈琼华罗聘婷
Owner GUANGZHOU UNIVERSITY
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