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Bacilli for environmental modification of stichopus japonicus aquaculture pond and application thereof

A bacillus and pond technology, which is applied in the field of screening beneficial microorganisms in aquatic products, can solve the problems of drug loss, environmental pollution, and hazards in adjacent waters, and achieve the effects of reducing residues, good growth, and strong ammonia nitrogen and COD degradation capabilities

Active Publication Date: 2014-08-27
MARINE BIOLOGY INST OF SHANDONG PROVINCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, it is unavoidable to use medicines for frequent diseases. Various antibiotics such as sulfonamides, tetracyclines, penicillins, furans, etc., result in secondary pollution on the breeding and produce many negative effects. For example, most of the invested medicines are directly lost to the environment. , directly pollute the environment and eventually pose a hazard to adjacent waters

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0050] Preparation of PCR template DNA:

[0051] Genomic DNA was extracted by boiling water. After the strain was activated, a single colony was picked and placed in 50 μl sterile distilled water, boiled in a boiling water bath at 100°C for 5-10 minutes, centrifuged and the supernatant was taken as PCR template DNA.

[0052] Determination of the 16SrDNA sequence of the strain:

[0053]16SrDNA amplification primers are bacterial 16SrDNA universal primers, forward primer: 5′-AGAGTT TGA TCC TGG CTC AG-3′(27F), reverse primer: 5′-TAC GGC TAC CTT GTT ACG ACT T-3′(1492R ). According to the PCR system (Niu Yufeng et al., 2009), the strains were amplified by PCR reaction. The PCR product (about 1.5kb) was detected by electrophoresis and sent to Beijing Sanbo Polygala Biotechnology Co., Ltd. for purification and sequencing. The sequencing results were compared with the 16S rRNA sequences of related genera and species in GenBank (http: / / www.ncbi.nlm.nih.gov) using Blast software, an...

Embodiment 1

[0060] Pick a single colony with an inoculation needle and inoculate them into corresponding sterilized conical flasks marked with A1, A2, A3, A4, A5, A6, A7 and A8 respectively, and the two conical flasks are not inoculated as blank. All the Erlenmeyer flasks were shaken at 28°C and 160r / min in a constant temperature shaker for 5 days. Samples were taken from the corresponding Erlenmeyer flasks and centrifuged at 5000r / min for 10min to get the supernatant, and the contents of COD and NH4-N in the supernatant were measured. Use 100×(1-COD5 / COD0)% and 100×(1-NH4-N5 / NH4-N0)% to represent the 5d degradation rates of each strain on organic pollutants and ammonia nitrogen in the A. japonicus bait. The specific values ​​are shown in the table below.

[0061] Strain number

COD degradation rate

Ammonia nitrogen degradation rate

A1

73.70％

67.10％

A2

22.80％

-82.60％

A3

9.80％

-6.70％

A4

3.60％

26.80％

A5

...

Embodiment 2

[0066] Prepare four 250mL Erlenmeyer flasks containing 100mL sterilized bait medium, use the inoculation needle to pick strain A1 and two existing bacillus (A8, A9) on the market, and inoculate them into three sterilized Erlenmeyer flasks respectively , an Erlenmeyer flask not inoculated as a blank. Place the Erlenmeyer flask in a constant temperature shaker at 17°C and 160r / min and shake it for 5 days. Corresponding Erlenmeyer flasks were sampled and centrifuged at 5000r / min for 10 minutes to get the supernatant, and the contents of COD and NH4-N in the supernatant were measured. Use 100×(1-COD5 / COD0)% and 100×(1-NH4-N5 / NH4-N0)% to represent the 5d degradation rates of each strain on organic pollutants and ammonia nitrogen in the A. japonicus bait. After 5 days, the COD and ammonia nitrogen degradation rates of strain A1 were 57.73% and 38.06%, respectively. The COD and ammonia nitrogen degradation rates of Bacillus A8 in the market were 49.05% and 18.31%, respectively, and ...

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Abstract

The invention provides the (bacilli sp.) A1 which is stored in a common microorganism center of the China Committee for Culture Collection of Microorganisms of Microbiology Research Institute of China Science Academy which is located at No.3 of yard 1 of the Beichen west road of Chaoyang District of Beijing, and the storage number is CGMCC No. 8686. The provided bacillus strain is original bacilli obtained by being directly separated and screened from aquatic water, and therefore the provided bacillus strain is low in potential fulminant perniciousness to stichopus japonicas and environment ecology when applied to a stichopus japonicus aquaculture pond. The bacillus strain is screened through low temperature and organic degradation capacity to obtain the target bacillus strain, and the target bacillus strain grows well under low temperature and is high in degradation capacity on ammonia nitrogen and COD in bait.

Description

technical field [0001] The invention belongs to the technical field of screening beneficial microorganisms in aquatic products, and in particular relates to a bacillus used for environmental restoration of sea cucumber culture ponds. Background technique [0002] The sea cucumber (Apostichopus japonicus Selenka) aquaculture industry is a new marine aquaculture industry. Since 2002, the sea cucumber aquaculture industry has developed rapidly. In 2006, the national sea cucumber aquaculture area reached 84,000 hectares, the output reached 76,000 tons, and the output value was close to 100 million. billion. However, with the rapid development of its breeding scale, related problems have gradually emerged. At present, most sea cucumber breeding mainly relies on artificial feed, which has poor adhesion, and a large number of artificial attachment bases are specially set up at the bottom of the pond, so a large amount of residual bait and feces will be produced during the breeding...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34A61K35/74A61P37/04A23K1/16A23K1/18C12R1/07A23K10/18A23K50/80
Inventor 宋爱环辛美丽刘元文邹琰王英俊孙福新
Owner MARINE BIOLOGY INST OF SHANDONG PROVINCE
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