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Safety evaluation method for edwardsiella disease phage therapy in aquaculture

A technology for safety evaluation and aquaculture, applied in the field of safety evaluation of Edwardsiella disease phage therapy in aquaculture, can solve the problems of lack of application, limited number of pure cultures, and lack of comprehensive understanding of the impact of fish safety. To achieve the effect of prevention and control

Pending Publication Date: 2022-07-26
UNIVERSITY OF CHINESE ACADEMY OF SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

On the one hand, due to the limited number of pure cultures of Edwardsiella phage obtained, on the other hand, because phage therapy has not received enough attention, its application in the prevention and control of Edwardsiella caused by Edwardsiella is still relatively scarce, and its impact on The impact on fish safety (such as antioxidant capacity and immune system) is even less fully understood

Method used

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  • Safety evaluation method for edwardsiella disease phage therapy in aquaculture
  • Safety evaluation method for edwardsiella disease phage therapy in aquaculture
  • Safety evaluation method for edwardsiella disease phage therapy in aquaculture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Comparison of zebrafish infection symptoms

[0022] Four groups of fish tanks were set up, namely control group (PBS+SM), pathogen infection group (E.p.+SM), antibiotic treatment group (E.p.+FLO) and phage treatment group (E.p.+EPP-1), with 30 fish in each group, It is administered by intraperitoneal injection, specifically as figure 1 shown. Except for the control group, the infection dose of the other groups was 10 5 CFU / tail fish, the concentration of florfenicol in the antibiotic treatment group was 10 mg / kg fish weight (the standard dose recommended by the FDA for the prevention and control of Edwardsiella), and the concentration of phage EPP-1 in the phage treatment group was MOI=1. Continuous monitoring was performed for 7 days. Mortality of zebrafish was recorded daily and survival curves were drawn. The zebrafish are fed with Artemia hatched by themselves in the laboratory. They are fed once a day at 9:00 am and 16:00 pm. The amount of Artemia is s...

Embodiment 2

[0024] Example 2 Comparison of therapeutic effects between bacteriophage EPP-1 and florfenicol

[0025] The survival curves of zebrafish under different treatment regimens are as follows: image 3 As shown, the zebrafish survival rate of the control group (PBS+SM) was 100% in the 7-day culture period, which means that the PBS and SM buffers are harmless to the zebrafish and the physical manipulation of the injection will not cause the zebrafish to die; In the pathogen infection group (E.p.+SM), the survival rate of zebrafish was only 13.33% in the 7-day culture cycle; in the antibiotic treatment group (E.p.+FLO), the zebrafish survival rate was 33.33% in the 7-day culture cycle. , compared with the pathogen infection group, it increased by 20%, reaching a very significant level (p<0.0001, Mantel-Cox test); the phage-treated group (E.p.+EPP-1) in the 7-day breeding cycle, the survival of zebrafish The rate was 30%, which was increased by 16.7% compared with the pathogen infect...

Embodiment 3

[0026] Example 3 Microscopic effects of bacteriophage EPP-1 and florfenicol on the body

[0027] Zebrafish were randomly removed on days 1, 3, and 7 of the culture cycle, and guts and livers were preserved for determination of antioxidant capacity and immune factors after dissection. The specific experimental setup was the same as that of the survival curve determination. Three zebrafish were randomly selected for each experiment as three biological replicates, and GSH, TNF-α and IL-6 in the intestine and liver of each fish were measured respectively. The zebrafish were dissected under a stereoscope, and the intestinal and liver tissues were dissected and placed in sterile centrifuge tubes, stored at -20°C, and the enzyme activities and immune factors were determined within 24 hours. The enzyme activity assay kit was purchased from Nanjing Jiancheng Biological Co., Ltd., and the assay method was carried out according to the kit instructions. Immune factors were determined by ...

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Abstract

The invention discloses a safety evaluation method for edwardsiella disease phage therapy in aquaculture, and belongs to the technical field of biology. The evaluation method disclosed by the invention is further applied on the basis of the obtained Edwardsiella bacteriophage EPP-1, and a safety evaluation method is provided for a bacteriophage prevention and control scheme of Edwardsiella disease frequently occurring in aquaculture. Compared with florfenicol, the two compounds have equivalent curative effects, so that the compound has a bright prospect when being used as an edwardsiella disease biological control agent in aquaculture; the enzyme activity and inflammatory cytokines in the intestinal tract and liver of the zebra fish are measured to characterize the influence of the enzyme activity and inflammatory cytokines on the antioxidant capacity and immune system of organisms; compared with florfenicol, the bacteriophage EPP-1 has the same curative effect, but has less influence on the oxidation resistance and immune system of zebra fish, and can be used as a potential biological agent for preventing and treating Edwardsiella disease in aquaculture.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for evaluating the safety of phage therapy for Edwardsiosis in aquaculture. Background technique [0002] Edwardsiellapiscicida (Edwardsiellapiscicida) belongs to the Enterobacteriaceae family, Edwardsiella genus, is a notorious fish pathogen that can cause serious Edwardsi economic losses. Once fish are infected with Edwardsiella pisiformis, severe immune dysfunction often occurs in multiple organs, resulting in organomegaly, ecchymosis, skin erosion, gillitis, ascites, and behavioral abnormalities typical of Edwardsiella. Disease clinical symptoms. The U.S. Food and Drug Administration (FDA) recommends taking 10-15 mg of florfenicol per kilogram of fish for no more than 10 consecutive days to control Edwardsiosis in aquaculture. But even environmental concentrations of antibiotic exposure can cause abnormal oxidative stress and immune dysregulation in different growth s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00
CPCA61K49/0004
Inventor 刘如铟韩岗华黄婷刘新春余志晟
Owner UNIVERSITY OF CHINESE ACADEMY OF SCIENCES
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