Method for separating heterotrophic ammonia-oxidizing microorganism from oligotrophic aquatic sediment

A technology for ammonia oxidizing microorganisms and nutrient-poor water bodies, which is applied in the field of environmental protection and can solve problems such as underestimation of microbial functions

Active Publication Date: 2014-05-07
GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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  • Description
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  • Application Information

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Problems solved by technology

[0003] Due to previous neglect of the heterotrophic ammonia oxidation process in nature, its microbial function has been greatly underestimated

Method used

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  • Method for separating heterotrophic ammonia-oxidizing microorganism from oligotrophic aquatic sediment
  • Method for separating heterotrophic ammonia-oxidizing microorganism from oligotrophic aquatic sediment

Examples

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Effect test

Embodiment 1

[0024] (1) Take 100g of sediment on the surface (5-10cm) of the bottom sediment of Xinfeng River in Heyuan, after conventional isolation, removal of sand and sedimentation, pre-aerate for 5 minutes, and put it into the sequence batch according to the mass ratio of mud to water at 1:1 In the first stage of the reactor, M0 liquid medium was added for enrichment, and the reactor was cultivated for 2 weeks. The residence time of the reactor in the first stage was 3-4 days. Enrichment of ammonia oxidizing microorganisms; 2. Screening of non-oligotrophic ammonia oxidizing microorganisms, the structure of the domestication device (sequencing batch reactor) is as follows figure 1 As shown, the culture temperature is 25°C, adjusted with sodium bicarbonate to maintain the pH between 6-7, without aeration, observe the clarification of the medium in the first stage of cultivation, and transfer to the second stage of cultivation after the sediment is flocculent. Gradually replace the M1 li...

Embodiment 2

[0035] (1) Take 300g of the bottom sediment surface (4-8cm) of the Dongjiang section of Heyuan, after conventional isolation and sedimentation, pre-aerate for 5 minutes, and put it into the sequencing batch reactor according to the mass ratio of mud to water at 1:1 for enrichment Set, add M0 liquid medium in the first stage of the reactor, and cultivate for 4 weeks. The residence time of the reactor in the first stage is 3 days. The culture medium was observed to be clarified in the first stage, and the sediment was flocculent and transferred to the second stage for cultivation. Then gradually replace the M1 liquid medium for batch culture. Cultivate for 6 weeks, the residence time of the second stage is 5-7 days, the culture temperature is 20°C, the pH is adjusted to be between 7-8 with sodium bicarbonate, no aeration is required, and the pre-culture solution containing domesticated flora is obtained through cultivation.

[0036] (2) Take 20ml of the bacteria solution from t...

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Abstract

The invention discloses a method for separating a heterotrophic ammonia-oxidizing microorganism from an oligotrophic aquatic sediment. The method comprises the steps: a, separating and settling a surface sediment of a bottom layer sediment in a clean river, and then, placing the surface sediment into a sequencing batch reactor for enriching culture to obtain a domestication flora containing preculture solution; b, taking liquid out of the preculture solution, and separating and purifying microorganisms in an enriched culture solution; c, carrying out a heterotrophic ammonia-oxidizing activity experiment: measuring ammonia nitrogen degradation activities of the enriching cultured microorganisms by using a special liquid culture medium, and selecting the microorganism with the ammonia nitrogen degradation efficiency of over 60% after being cultured for 2-3 days to obtain an efficient strain, wherein the enriching cultured microorganisms are separated and purified in the step b, and d, separating, purifying and carrying out enriching culturing on the obtained efficient strain in the step c to obtain the heterotrophic ammonia-oxidizing microorganism.

Description

Technical field: [0001] The invention belongs to the technical field of environmental protection, and in particular relates to a method for separating heterotrophic ammonia oxidizing microorganisms from nutrient-poor water sediments. Background technique: [0002] Heterotrophic ammonia oxidizing microorganisms are a new group of ammonia oxidizing microorganisms that have been widely concerned in recent years. The basic principle of heterotrophic ammonia oxidation is that such microorganisms simultaneously convert ammonia nitrogen and organic matter into nitrogen and carbon dioxide under aerobic conditions, so as to simultaneously degrade rivers. The function of ammonia nitrogen and organic matter in the medium. The nitrogen cycle is one of the main pathways of the earth's element cycle. When nitrogen-containing pollutants are decomposed, they will enter the first core link of the nitrogen cycle: the ammonia oxidation process. However, the autotrophic ammonia oxidation proce...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02C12R1/01
Inventor 钟玉鸣许玫英郭俊孙国萍潘国平连英丽
Owner GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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