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Probiotic separated from intestinal tracts and application thereof

A probiotics, intestinal technology, applied in applications, medical preparations containing active ingredients, bacteria, etc., can solve problems such as drug resistance and drug residues, endangering human health, etc., achieve good application prospects, improve survival rate and anti- The effect of sickness

Active Publication Date: 2014-01-22
YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, chemical therapy is still widely used in the treatment of aquatic animal diseases. The abuse of antibiotics makes water bacteria resistant to drugs and drug residues, which indirectly endangers human health.

Method used

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  • Probiotic separated from intestinal tracts and application thereof
  • Probiotic separated from intestinal tracts and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Screening and separation of embodiment 1 probiotics

[0017] 1) Sampling: 5-10 healthy turbot (body length 10-15cm) were randomly selected, anesthetized with 100-200ul of 1-2% pentobarbital sodium, dissected under sterile conditions, and their intestines were taken. Quickly wash with sterile 2216E liquid medium (ingredients: peptone 5g, yeast extract 1g, ferric phosphate 0.1g, aged sea water 1000mL, pH7.5, sterilized at 121°C for 20min) or sterile PBS, 5-10mL glass homogenate Homogenate, homogenate with sterile 2216E liquid medium or PBS according to 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 Gradient dilution. choose 10 -3 and 10 -6 The diluted homogenate was uniformly coated on TSB, LB, and 2216E solid plates. 28-30°C, culture upside down for 24-48h.

[0018] 2) Preliminary screening: the single colony grown on the above-mentioned plate is picked up according to the morphological and structural characteristics of the colony, and cultivated to OD with the liqui...

Embodiment 2

[0022] Example 2: Identification and activity analysis of P115

[0023] 1.16S rDNA analysis

[0024] a) Extraction of P115 genomic DNA: culture P115 to OD 600 =1.0-1.5, use the phenol-chloroform method to extract the genomic DNA of P115 to produce a protein-free layer, and dissolve it with a certain amount of TE solution.

[0025] b) PCR amplification of 16SrDNA: PCR reaction system: 0.5 μl P115 genomic DNA, 1 μl 27F, 1 μl 1492R, 4 μl dNTP, 5 μl Taqbuf, 0.4 μl Taq enzyme, 36.1 μl ddH 2 O. The reaction conditions were pre-denaturation at 95°C for 5 min, denaturation at 95°C for 1 min, renaturation at 54°C for 30 s, extension at 72°C for 1 min, and 30 cycles. Extend at 72°C for 10 min. The PCR product was detected as a single band by electrophoresis, with a size of about 1.6 kb, which was in line with expectations. Among them, specific primers 27F and 1492R are commonly used primers and belong to the public domain.

[0026]

[0027] c) Sequencing and analysis: After reco...

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Abstract

The invention relates to the field of microbial disease control and in particular relates to a probiotic separated from intestinal tracts of turbots and an application thereof. The probiotic is Leclercia adecarboxylata P115, is collected in China General Microbiological Culture Collection Center (CGMCC) on June 24, 2013, and has a collection number of CGMCCNo.7813. The probiotic is separated from the intestinal tracts of healthy turbots. The probiotic and the pure culture thereof have obvious bacteriostatic effects on tritirachium album, klebsiella pneumoniae, staphylococcus aureus, vibrio parahaemolyticus, vibrio harveyi and vibrio anguillarum. The pure culture or composition of the probiotic can be used for control of aquaculture diseases and development of novel antibacterial materials, and has good application prospects.

Description

technical field [0001] The invention relates to the field of microbial disease control, in particular to a probiotic isolated from the intestinal tract and its application. Background technique [0002] Probiotics are a class of active microorganisms that are beneficial to the host. They are a general term for active beneficial microorganisms that colonize the intestinal tract and reproductive system of humans or animals and can produce definite health effects to improve the host's micro-ecological balance and exert beneficial effects. It works primarily by improving the balance of the host's gut microbiota. It has the functions of improving the structure of intestinal flora, inhibiting pathogenic bacteria, generating nutrients, improving the body's immunity, eliminating carcinogenic factors, lowering cholesterol and blood pressure, and improving lactose digestibility. Fuller first defined probiotics in 1989: "Probiotics are live bacterial additives that have a beneficial e...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23K1/16A61K35/74A61P31/04A61P31/10C12R1/01A61K35/741
Inventor 焦绪栋秦松
Owner YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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