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Aerobic denitrification preparation and its preparation method and application

A preparation and oxygen removal technology, applied in the field of environmental microorganisms, can solve the problems of water quality and ecological balance impact or interference, and achieve the effect of significant immune regulation.

Active Publication Date: 2022-03-29
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, this type of external carbon source itself has no selectivity and can be utilized by any microorganisms including pathogenic microorganisms
Water quality and ecological balance may therefore be affected or disturbed

Method used

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  • Aerobic denitrification preparation and its preparation method and application
  • Aerobic denitrification preparation and its preparation method and application
  • Aerobic denitrification preparation and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] The screening of embodiment 1 PHAs degrading bacterial strain AOB-7

[0048] The PHAs degrading strain AOB-7 was obtained by screening in DM-PHAs medium. details as follows:

[0049] Prepare DM-PHAs medium with PHAs. The composition of DM-PHAs per liter solution is as follows: KNO 3 1.00g, PHAs particles 3.0g, MgSO 4 ·7H 2 O 0.20g, CaCl 2 0.01g, KH 2 PO 4 0.50g, Na 2 HPO 4 0.50g, FeSO 4 0.01g, NaCl10.00g, trace elements 1.00mL, pH 7.2. The composition of the above trace elements is as follows: 57.10g / L EDTA·2Na, 3.90g / LZnSO 4 ·7H 2 O, 7.00g / L CaCl 2 2H 2 O, 1.00g / L MnCl 2 4H 2 O, 5.00g / L FeSO 4 ·7H 2 O, 1.10g / L (NH4) 6 Mo 7 o 24 4H 2 O, 1.60g / L CuSO 4 ·5H 2 O, 1.60g / L CoCl 2 ·6H 2 O, pH 6.0.

[0050] Activate and cultivate the strain to be screened in LB liquid medium, culture it on a shaker at 30°C and 150rpm for 24 hours, centrifuge at 8000g for 10 minutes, discard the supernatant, wash the bacteria three times with 0.9% normal saline, a...

Embodiment 2

[0054] Example 2 Detection of the aerobic denitrification characteristics of the PHAs degrading strain AOB-7 using PHB and PHBV as the only carbon source

[0055] Inoculate the PHAs-degrading strain AOB-7 into DM-PHB or DM-PHBV medium, culture on a shaker at 25-30°C and 120-150rpm for 84-108 hours, and detect the OD of the bacteria in the culture system every 12 hours 600 Value, nitrite nitrogen, nitrate nitrogen concentration changes, according to whether nitrate is reduced to nitrite, determine the characteristics of its aerobic denitrification denitrification.

[0056] The growth of strain AOB-7, the concentration changes of nitrite nitrogen and nitrate nitrogen are as follows: figure 1 As shown, after cultured in DM-PHB and DM-PHBV medium for 4 days 600 The maximum values ​​are reached, 4.2 and 2.6, respectively. Further verification of OD by plate colony counting 600 value. The total colony forming units in DM-PHB and DM-PHBV were 1.41 × 10 9 CFU / mL and 3.23×10 8 CF...

Embodiment 3

[0061] Example 3 Detection of gaseous products produced by PHAs degrading strain AOB-7 using PHB and PHBV during aerobic denitrification

[0062] The above aerobic denitrification process shows that strain AOB-7 can carry out efficient aerobic denitrification pathway, and NO 3 - -N or NO 2 - -N is converted to gaseous nitrogen. To test this speculation, GC-MS and GC-IRMS analyzes were performed to identify the N produced by the aerobic denitrification pathway 2 O and N 2 .

[0063] For the gaseous product N of the denitrification process 2 and N 2 The detection of O is the most conclusive and direct evidence to determine whether the strain has denitrification function. Due to the presence of a large amount of N in the air 2 , the N produced by the denitrification pathway 2 The amount is small, simply by detecting N 2 The change of the amount does not explain the problem, so combined with isotope labeling, it can be verified by detecting the change of isotope ratio i...

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Abstract

The invention provides an aerobic denitrification preparation, a preparation method and application thereof. The aerobic denitrification preparation is fermented by inserting Pseudomonas AOB-7 into a medium containing biodegradable materials, and the obtained fermentation product is used as an aerobic denitrification preparation. Strain AOB‑7 is a Pseudomonas strain that can use PHAs as a solid slow-release carbon source and carrier for efficient aerobic denitrification. Based on the strain AOB‑7, a PHA particle was developed as a solid slow-release carbon source and biofilm carrier A new type of aerobic denitrification preparation, this new type of aerobic denitrification preparation can be directly added to low carbon nitrogen ratio water bodies (such as aquaculture ponds) to improve water quality, and gaseous nitrogen products can be produced during the aerobic denitrification process. The nitrate nitrogen accumulated in the water body is converted into gas to achieve complete denitrification. In addition, this new aerobic denitrification preparation can produce 3‑hydroxybutyric acid, which has immunomodulatory effects on aquaculture animals, during the aerobic denitrification process.

Description

technical field [0001] The invention belongs to the technical field of environmental microorganisms, and in particular relates to an aerobic denitrification preparation and its preparation method and application, especially the application in denitrification of aquaculture circulating water. Background technique [0002] In recent years, with the development of the economy and the improvement of people's living standards, high-density aquaculture has flourished all over the world. Recirculating aquaculture system (RAS) has been widely used at home and abroad because of its advantages of high intensification, environmental friendliness, resource saving, controllable farming, and high yield. Ammonia nitrogen is the most important metabolic waste produced by aquaculture animals. When ammonia nitrogen exists in the state of molecular ammonia, it will cause strong neurotoxicity to aquaculture animals. Acute ammonia poisoning is one of the most serious hazards in aquaculture water...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C02F3/02C02F3/34C12P7/42C12R1/38C02F101/16
CPCC02F3/02C02F3/34C12P7/42C02F2101/163C02F2101/166C12R2001/38C12N1/205Y02W10/10
Inventor 刘志培高喜燕刘缨苗莉莉
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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