Efficient water body denitrification pseudomonas stutzeri DB-33 and culture method thereof

A technology of Pseudomonas stutzeri and DB-33, which is applied in the direction of chemical instruments and methods, methods based on microorganisms, biochemical equipment and methods, etc., to achieve good degradation effects, increase the simplicity of production operations, and reduce production costs Effect

Inactive Publication Date: 2012-09-19
TIANJIN CITY AGRI BIO TECH RES CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to overcome the defects of the existing biological denitrification technology of polluted water bodies, aiming to develop a kind of high-efficiency denitrification bacteria and its simple

Method used

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  • Efficient water body denitrification pseudomonas stutzeri DB-33 and culture method thereof
  • Efficient water body denitrification pseudomonas stutzeri DB-33 and culture method thereof
  • Efficient water body denitrification pseudomonas stutzeri DB-33 and culture method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Isolation, screening and identification of strains

[0040] (1) Strain isolation:

[0041] Sample collection: In October 2007, samples of bottom sediment from fish and shrimp breeding ponds in Xiqing District, Tianjin were collected.

[0042] Sample enrichment: configure liquid enrichment medium:

[0043] DM medium: sodium citrate 5 g, KNO 3 2 g, KH 2 PO 4 1 g, MgSO 4 ·7H2O 0.2 g, distilled water 1 000 ml, pH7.2

[0044] Take 3-5 g of the sediment sample and add it to the enrichment medium, and incubate at 30°C for 5 days.

[0045] Separation medium

[0046] BTB (thymol blue) solid medium: asparagine 1 g, KNO 3 1 g, KH 2 PO 4 1 g, FeC1 2 ·6H 2 O 0.5 g, CaCl 2 -2H 2 O 0.2 g, MgSO 4 ·7H 2 O 1 g, sodium succinate 8.5 g, agar 15-18 g, 1% bromothymol blue (dissolved in absolute ethanol) 1 mL, distilled water 1 000 mL, pH 7-7.5

[0047] The enriched bacterial liquid was streaked and separated on the separation medium, and a single strain was obtained afte...

Embodiment 2

[0059] Nitrogen removal effect of DB-33 strain in simulated aquaculture wastewater

[0060] Degradation Effect of DB-33 Strain on Nitrate

[0061] Fill the glass container with 2.5 L of simulated aquaculture wastewater, and adjust the initial concentration of nitrate nitrogen to 138 mg L -1 , put 50 mL of bacterial solution into it and place it in a 30°C incubator for cultivation. Taking no bacteria as a control, samples were taken regularly to measure the nitrate content in the water. The result is as figure 1 .

[0062] Depend on figure 1 It can be seen that when denitrifying bacteria were introduced into the water body, the nitrate concentration within 48 hours was 138mg L -1 The degradation rate of the simulated wastewater was 94.58%, and the nitrite was completely degraded within 72 hours, and the degradation effect was significantly better than that of the control.

[0063] Degradation Effect of DB-33 Strain on Nitrite

[0064] Fill the glass container with 2.5 L...

Embodiment 3

[0070] Fermentation of DB-33 strain under open conditions

[0071] Activation: Prepare DM solid medium, sterilize it and pour it into a 90mm Petri dish. After cooling, insert the preserved DB-33 slant strain and incubate at 30°C for 24h.

[0072] Production of liquid strains: Prepare DM liquid medium, divide into Erlenmeyer flasks, sterilize, and after cooling, pick activated strains from petri dishes and incubate at 30°C for 24 hours.

[0073] Plastic barrel aerated fermentation: Use a 25L plastic barrel as a culture device. The old barrel is sterilized with 1% sodium hypochlorite solution, and then rinsed with clean water. The new barrel can be washed with clean water and used directly. Load 20L of DM liquid culture medium into the liquid bacteria in the triangular flask, the inoculated amount is 20%, the temperature is above 25°C, and the air pump (45w) is connected to the sand head (soaked in 2% sodium hypochlorite solution) for ventilation. The fermentation time is 24 ho...

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Abstract

The invention discloses a pseudomonas stutzeri DB-33 used for the quick denitrification of a culture water body, which has a collection number of CGMCC (China General Microbiological Culture Collection Center) No. 5993. The invention also discloses an open culture method for the fungicide, which comprises the following steps of: taking a 25L plastic drum as a culture device; filling in 20L of DM liquid culture medium; inoculating a liquid strain with the strain inoculation amount of 20%; at the temperature above 25DEG C, ventilating for 45w by an air pump; and fermenting for 24 hours until the effective viable count of the fermentation liquor reaches 16 hundred million mL<-1>. The pseudomonas stutzeri disclosed by the invention can be used for quickly degrading nitrate, nitrite and ammonia nitrogen in the water body so as to improve the water body environment, reduce harm to organisms in the water body by nitrogen-contained pollutants and greatly lower the production cost of a denitrification microorganism preparation, and the efficient, safe and practical biological denitrification technology is provided for vast farmers.

Description

technical field [0001] The invention belongs to the technical field of biological bacteria preparation and the application field of environmental microorganisms. The invention relates to a Pseudomonas stutzeri DB-33 applied to rapid denitrification of aquaculture water and an open culture method thereof. Background technique [0002] According to the State Environmental Protection Administration’s 2010 Bulletin on the State of the Environment in China, the main lakes in my country are heavily polluted with nitrogen and phosphorus. Due to the increase in breeding density and excessive feed input, the problem of eutrophication in aquaculture water is prominent. High concentrations of ammonia nitrogen and (sub)nitrate nitrogen not only have a direct toxic effect on many aquatic animals, but also reduce their immunity, resulting in frequent occurrence of aquatic diseases and bringing huge losses to the aquaculture industry. Therefore, to solve the problem of eutrophication of w...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C12R1/38C02F101/16
Inventor 张峰峰赵玉洁谢凤行周可
Owner TIANJIN CITY AGRI BIO TECH RES CENT
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