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Lactobacillus casei YFI-5 and application thereof in resistance of cyprinid herpesvirus 2

A technology of Lactobacillus casei and carp herpes virus, applied in the direction of Lactobacillus, antiviral agent, application, etc., to prevent and treat hematopoietic organ necrosis of crucian carp, easy to use, and reduce mortality

Active Publication Date: 2019-11-19
YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, lactic acid bacteria are widely used in aquaculture, but there are few studies on lactic acid bacteria inhibiting aquatic pathogens and viruses

Method used

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  • Lactobacillus casei YFI-5 and application thereof in resistance of cyprinid herpesvirus 2
  • Lactobacillus casei YFI-5 and application thereof in resistance of cyprinid herpesvirus 2
  • Lactobacillus casei YFI-5 and application thereof in resistance of cyprinid herpesvirus 2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Isolation and identification of strain YFI-5

[0025] 1. Isolation and identification of strain YFI-5

[0026] Lactobacillus casei was isolated from pond water samples in aquaculture areas. Specifically, 0.85% sterile saline was used to dilute the culture pond water samples 10 times in a row for 6 times, and the samples were diluted in each concentration gradient dilution with a pipette gun. Draw 100 μL of the solution onto the BHI solid plate, spread it evenly with a spreader, number it, and do 3 repetitions. After coating evenly, place it in the ultra-clean workbench for 5-10 minutes, so that the bacterial liquid on the surface of the medium is fully absorbed. Finally, the plate was inverted and incubated in a constant temperature incubator at 30°C for 24h. Bacterial colonies of different forms were selected and inoculated on ordinary broth plates for separation and purification, and the antiviral function of different bacteria was tested, and finally a strain resis...

Embodiment 2

[0035] Embodiment 2: antiviral spectrum detection of Lactobacillus casei YFI-5 fermentation supernatant

[0036] Pick a single colony of rejuvenated Lactobacillus casei YFI-5 in 150ml MRS liquid medium, incubate at 37°C for 24h, centrifuge at 5000rpm for 10min, filter the supernatant with a 0.22μm filter membrane, and store it in a sterile centrifuge tube4 Store at ℃ for later use. The effects of fermentation supernatant of Lactobacillus casei YFI-5 on carp herpesvirus type II (CyHV-2), giant salamander iridescent virus (GSIV), grass carp reovirus (GCRV) and carp spring viremia virus (SVCV) were detected respectively. Inhibitory effect.

[0037] The cells used in this example are crucian carp brain tissue cell line (GiCB, CCTCC NO: C2013179), giant salamander muscle cell line (GSM), grass carp kidney cell line (CIK), carp epithelial cell line (EPC).

[0038] The volume of the fermentation supernatant of Lactobacillus casei YFI-5 added in this embodiment is 100 μ L per hole, ...

Embodiment 3

[0043] Application of Lactobacillus casei YFI-5 fermentation supernatant in anti-carp herpesvirus type II (CyHV-2):

[0044] Pick a single colony of rejuvenated Lactobacillus casei YFI-5 in 150ml MRS liquid medium, incubate at 37°C for 24h, centrifuge at 5000rpm for 10min, filter the supernatant with a 0.22μm filter membrane, and store it in a sterile centrifuge tube4 ℃ for future use, and used in this example and Example 4.

[0045] The cells used in this example are crucian carp brain tissue cell line (GiCB), CCTCC NO: C2013179. The cell maintenance solution used is M199 medium with 10% FBS;

[0046] The volume of the fermentation supernatant of Lactobacillus casei YFI-5 added in this embodiment is 100 μ L per hole, and the added 100 TCID 50 / 0.1ml of the CyHV-2 virus liquid has the same volume as the fermentation supernatant;

[0047] Detection of virus titer: adding 1×10 4 100 μL of GiCB cells per well, cultured at 25°C for 16-24 hours. When the cells grow to 80%-90%,...

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Abstract

The invention relates to the technical field of aquatic antiviral microecological preparation, and in particular, relates to lactobacillus casei YFI-5 and an application thereof in resistance of cyprinid herpesvirus 2 (CyHV-2). The lactobacillus casei has the preservation number of CCTCC NO:M2019654. A fermentation supernatant of the lactobacillus casei YFI-5 can reduce the infection activity of CyHV-2 on host cells and inhibit the infection of CyHV-2 virus invasion on the cells, and has no side effects on cells. The feed with addition of the lactobacillus casei YFI-5 can effectively reduce the mortality rate of crucian carp infected by CyHV-2, and can be used for preventing and treating crucian carp hematopoietic necrosis.

Description

technical field [0001] The invention relates to the technical field of aquatic antiviral microecological preparations, in particular to a kind of Lactobacillus casei YFI-5 and its application in anti-carp herpesvirus type 2. Background technique [0002] Crucian carp is one of the important species of freshwater aquaculture fish in my country, and its annual output has exceeded 3 million tons, occupying a very important position in freshwater aquaculture. In recent years, the hemorrhagic disease of crucian carp in my country has occurred successively, leading to fulminant death of cultured heterogeneous gibel carp, causing heavy economic losses. Ultrathin slices of spleen and kidney tissues of sick crucian carp were observed by electron microscope, and a large number of typical herpes virus-like particles were found. Further research confirmed that the disease was Crucian Carp Hematopoietic Necrosis, and the pathogen was Cyprinid herpesvirus type II (Cyprinid herpesvirus2, ...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/747A61P31/22A23K10/18A23K50/80C12R1/245
CPCA61K35/747A61P31/22A23K10/18A23K50/80C12R2001/245C12N1/205A23V2400/125
Inventor 周勇薛明洋范玉顶孟彦刘文枝江南李逸群曾令兵
Owner YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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