A strain of Candida rugosa for heterotrophic nitrification and aerobic denitrification and its application

A technology of aerobic denitrification and heterotrophic nitrification, applied in the field of Candida rugosa strains of heterotrophic nitrification and aerobic denitrification

Active Publication Date: 2021-04-27
HUNAN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The nitrogen degradation function of Candida rugosa and its application to aquaculture sewage treatment have not been reported in domestic and foreign literature

Method used

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  • A strain of Candida rugosa for heterotrophic nitrification and aerobic denitrification and its application
  • A strain of Candida rugosa for heterotrophic nitrification and aerobic denitrification and its application
  • A strain of Candida rugosa for heterotrophic nitrification and aerobic denitrification and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Strain screening method

[0048] 1. Primary screening

[0049] Collect 10mL of water samples from freshwater aquaculture water with high ammonia nitrogen content, add them to 100mL culture medium containing 52.39mg / L ammonia nitrogen, enrich and cultivate at 30°C and 180r / min for 3 days, and obtain the enriched solution for later use.

[0050] 2. Dilute the prepared enrichment solution to 10 times according to the 10-fold dilution method -7 , take 10 respectively -5 、10 -6 、10 -7 Three dilutions of different dilutions were spread on a nitrated solid medium plate, and placed in a 30° C. incubator for 48 hours to obtain colonies.

[0051] Described nitrifying solid medium (g / L) formula and preparation method are as follows: (NH 4 ) 2 SO 4 ·7H 2 O 0.247g, K 2 HPO 4 7g, KH 2 PO 4 3g, MgSO 4 ·7H 2 O 0.1g, FeSO 4 ·7H 2 O 0.05g, H 2 O 1L, agar 20g. Dilute to volume with distilled water, the initial pH value is 7.0, and sterilize at 121°C for 20 minutes to mak...

Embodiment 2

[0063] Identification of Morphological and Physiological and Biochemical Characteristics of Strain DW-1

[0064] 1. Identification of Morphological Characters

[0065] The colony morphological characteristics of strain DW-1 are as follows:

[0066] The strain DW-1 was inoculated in the liquid seed medium, shaken at 30°C and 180r / min for 24 hours, and diluted to 10 according to the 10-fold dilution method. -8 , take 10 -6 、10 -7 、10 -80.1 mL of three different dilutions of dilutions were evenly spread on the surface of the PDA solid medium plate, and cultured at a constant temperature of 30°C for 24 hours. The characteristics of the observed colonies were as follows: the colonies were round, milky white, moist, smooth and opaque (see figure 1 and figure 2 ).

Embodiment 3

[0068] 26S rDNA sequence analysis

[0069] 1. Extraction of strain DW-1 genome

[0070] (1) Bacteria culture: Inoculate the strain DW-1 in PD liquid medium, shake culture at 30°C, 180r / min for 24 hours, and collect the bacteria by centrifugation at 4°C, 10000r / min.

[0071] (2) Genomic DNA extraction: The 26S rDNA D1 / D2 region genomic DNA of the DW-1 strain was extracted using the Ezup Column Yeast Genomic DNA Extraction Kit produced by Sangon Bioengineering (Shanghai) Co., Ltd.

[0072] (3) Preservation: the extracted DW-1 strain genomic DNA was stored at -20°C.

[0073] (4) Detection: After the extracted DW-1 strain genomic DNA was subjected to 1% agarose gel electrophoresis, it was observed and photographed in a gel imaging system, and the electrophoresis image was obtained as follows: image 3 shown.

[0074] 2. PCR amplification

[0075] The PCR amplification primers used universal primers NL1 (5'-GCATATCAATAAGCGGAGGAAAAG-3', SEQ ID No.1) and NL4 (5'-GGTCCGTGTTTCAAGAC...

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Abstract

The invention provides a heterotrophic nitrification-aerobic denitrification Candida rugosa strain and its application, belonging to the technical field of functional microorganisms. Heterotrophic nitrification and aerobic denitrification Candida rugosa (Diutina rugosa) strain DW‑1, the deposit number is CCTCC NO: M2019166. The invention also provides a microbial preparation for degrading ammonia nitrogen and nitrite nitrogen in freshwater aquaculture sewage comprising the bacterial strain DW‑1. DW-1 strain has the function of heterotrophic nitrification-aerobic denitrification, and can directly convert most of the ammonia nitrogen in nitrogen-containing sewage into gas products through heterotrophic nitrification-aerobic denitrification, and the accumulation of nitrite and nitrate nitrogen less; ammonia nitrogen or nitrite nitrogen can be used as the only nitrogen source for metabolism to achieve high-efficiency denitrification, so the application of the Candida rugosa strain DW-1 or the microbial preparation in the denitrification of freshwater aquaculture water.

Description

technical field [0001] The invention belongs to the technical field of functional microorganisms, in particular to a candida rugosa strain for heterotrophic nitrification and aerobic denitrification and application thereof. Background technique [0002] With the increasingly serious problem of eutrophication in freshwater aquaculture water, many large water surface seine aquaculture and fence aquaculture have been cancelled, and the implementation of policies such as "returning fish to lakes" has prompted freshwater aquaculture to play an irreplaceable role in my country's aquaculture industry. . However, with the increasing level of aquaculture and the growth of fish carrying capacity per unit of water, a large amount of feed and fish metabolites are put into the aquaculture water, causing serious pollution to the aquaculture water quality, and the aquaculture wastewater is directly discharged into the surrounding rivers without treatment, resulting in eutrophication of the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C02F3/34C02F101/16C12R1/72
CPCC02F3/347C02F2101/16C02F2101/166C12N1/14C12N1/165C12R2001/72
Inventor 兰时乐杜全能胡超朱文娟陈思宇肖思远
Owner HUNAN AGRICULTURAL UNIV
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