Probiotic with bacteriostatic ability and application thereof

A technology of probiotics and ability, applied in the field of probiotics, can solve problems such as drug resistance, drug residues, and harm to human health, and achieve good application prospects

Active Publication Date: 2014-01-22
YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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  • 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 ant

Method used

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  • Probiotic with bacteriostatic ability and application thereof
  • Probiotic with bacteriostatic ability and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Screening and isolation of probiotic Enterobacter ludwigii PC88

[0025] 1) Sampling: 5-10 healthy flounder (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 use sterile TSB liquid medium (ingredients: tryptone 17g, polyvalent peptone 3g, yeast extract 6g, sodium chloride 5g, dipotassium hydrogen phosphate 2.5g, glucose 2.5g, add water to 1L, pH=7.2-7.5) Or wash with sterile PBS, homogenize with 5-10mL glass homogenizer, use sterile TSB 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 evenly spread on TSB and LB solid plates respectively. 28-30°C, culture upside down for 24-48h.

[0026] 2) Preliminary screening: the single colony grown on the above plate, according to the morphological and structural characteristics of the colo...

Embodiment 2

[0030] Example 2: Identification and activity analysis of PC88

[0031] 1) 16S rDNA analysis

[0032] a) Extraction of PC88 genomic DNA: culture PC88 to OD 600 =1.0-1.5, use the phenol-chloroform method to extract the genomic DNA of PC88 to produce a protein-free layer, and dissolve it with a certain amount of TE solution.

[0033] b) PCR amplification of 16SrDNA: PCR reaction system: 0.5 μl PC88 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.

[0034] c) Sequencing and analysis: After recovering the gel of the PCR product, send it to a sequencing company for sequencing. Results After nBlast analysis,...

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Abstract

The invention relates to the field of microbial disease control and in particular relates to a probiotic with bacteriostatic ability. The probiotic is Enterobacter ludwigii-PC88, is collected in China General Microbiological Culture Collection Center (CGMCC) on June 24, 2013, and has a collection number of CGMCC No.7814. The probiotic is separated from intestinal tracts of healthy flounders. The pure culture of the probiotic has relatively strong inhibitory effects on aquatic pathogens such as vibrio splendidus, vibrio harveyi, vibrio parahaemolyticus, pseudomonas aeruginosa and vibrio anguillarum, also can inhibit growth of known human pathogens such as tritirachium album, staphylococcus aureus and klebsiella pneumoniae, and has resistance to ampicillin. 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 with antibacterial ability. 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 effect on the health of the h...

Claims

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Application Information

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