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Compound bacterial agent for nitrogen reduction, and application and preparing method thereof

A technology for compound bacteria and nitrogen reduction, which is applied in microorganism-based methods, biochemical equipment and methods, chemical instruments and methods, etc., can solve the high labor cost of engineering methods, the ecological risk of secondary pollution by chemical methods, and the effectiveness of physical methods. Slow and other problems, to achieve the effect of strong adaptability, easy cultivation and fast growth

Inactive Publication Date: 2019-09-03
陈海清
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among these methods, the chemical method is prone to secondary pollution and ecological risks, the physical method is slow to take effect, and the labor cost of the engineering method is too high; while the biological method is recognized as one of the most economical and effective methods for denitrification and nitrogen removal, among which widely used microbiological methods

Method used

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  • Compound bacterial agent for nitrogen reduction, and application and preparing method thereof
  • Compound bacterial agent for nitrogen reduction, and application and preparing method thereof
  • Compound bacterial agent for nitrogen reduction, and application and preparing method thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0039] The preparation process of the preparation includes compounding the separate cultures of the three bacterial strains and determining the compound preparation with the best effect; the compound preparation with the best effect includes No. Bacillus sp., Bacillus strain B No. Bacillus sp., Bacillus strain C No. Bacillus amyloliquefaciens the combination of three strain cultures. That is, the combination method of A# strain + B# strain + C# strain has the best effect and the best cost performance.

[0040] The present invention also provides a method for preparing a nitrogen-reducing preparation from the compound bacterial agent used for nitrogen reduction, comprising: step 1, strain isolation and cultivation; step 2, strain identification; step 3, fermentation and cultivation of strains, collection of fermentation products, and obtaining A single-ingredient preparation; then compound the three strains to obtain a compound preparation with the best effect.

[0041] Step 1...

Embodiment 1

[0056] Example 1 Bacterial classification isolation culture and bacterial strain identification.

[0057] 1. Isolation and cultivation of bacterial strains.

[0058] Pond sediment samples were selected for the isolation and screening of bacteria. After the collected samples were aseptically stored and brought back to the laboratory (Shenzhen Pintai Biotechnology Co., Ltd.), take 1g of the mud sample and add 100mL of sterile water, shake and mix well, after standing for a while, take the supernatant for doubling dilution, and dilute the diluted The solution was aseptically coated in the newly prepared LB solid medium, and cultured in a constant temperature incubator at 30°C for 24-48 hours. The monoclonal bacteria with obvious morphology were picked out and inoculated in the newly prepared LB liquid medium.

[0059] 2. Screening of nitrogen-reducing strains.

[0060] Cultivate the picked single clone until the bacterial concentration reaches OD 600 When = 0.6-0.8, add in the...

Embodiment 2

[0115] Example 2. Determination of inorganic nitrogen in the screening of nitrogen-reducing bacterial strains.

[0116] Specifically include the following processes:

[0117] ① Collection and treatment of urban domestic wastewater. The recovered wastewater samples are first filtered once with a 0.45 μm macroporous water phase filter membrane to remove particles and some microorganisms; then filtered once with a 0.22 μm water phase filter membrane to further remove tiny bacteria. The filtered urban domestic sewage is divided into 50mL sterilized culture tubes, with a liquid volume of 30ml, for use.

[0118] ②Preparation of nitrogen-reducing strains. The strains were cultured in test tubes (50mL total volume, liquid volume 20mL), and the OD of the four strains was detected by a multi-functional microplate reader (infinite M200PRO) during the culture 600 value, when the strain OD 600 When the value reaches 0.6-0.8, the strain can be used for subsequent experiments. Put 0.5ml...

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Abstract

The invention discloses a compound bacterial agent for nitrogen reduction, and an application and a preparing method thereof. The bacterial agent comprises three bacterial strains, all of the bacterial strains are gram-positive bacteria, namely bacillus sp. No.A, bacillus sp. No.B, and bacillus sp. No.C bacillus amyloliquefaciens. The invention also provides the application of the compound bacterial agent, wherein the bacterial agent is used for preparing a preparation for reducing inorganic nitrogen value in domestic wastewater. The invention also provides a method for preparing a nitrogen reducing agent with the compound bacterial agent. According to the compound bacterial agent, and the application and the preparation method thereof provided by the invention, the bacterial strain is from a natural environment, and has the advantages of no secondary pollution to the ecological environment, no ecological risk, strong adaptability, fast growth speed, easy cultivation and the like. Thecombination of the compound way obtains better nitrogen reduction effect than a single bacterial agent.

Description

technical field [0001] The invention relates to a compound bacterial agent comprising three denitrifying bacteria that reduce the inorganic nitrogen value in domestic sewage and its application and preparation method. Three nitrogen sources (NO 3 - , NO 2 - , and NH 4 + ) related bacillus (Bacillus sp.) compound bacterial agent and its application and preparation method. Background technique [0002] In recent years, with the increasing urban population and the rapid development of industrialization, the discharge of urban sewage and industrial wastewater in my country has become increasingly serious, leading to the deterioration of water quality and environment, the increase of eutrophication and the decrease of biodiversity, etc., which seriously threaten the ecology of water bodies. Health and ecological civilization construction. In the detection of water quality parameters, nitrogen (N) element is an important indicator of water quality load, and the excess of nit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02C02F3/34C12R1/07C02F101/16
CPCC02F3/34C02F2101/163C12N1/02C12N1/20
Inventor 陈海清
Owner 陈海清
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