Tilapia mossambica streptococcus agalactiae oral attenuated freeze-dried vaccine

A technology of streptococcus lactis and tilapia, which is applied in the direction of freeze-dried transportation, antibacterial drugs, bacterial antigen components, etc., can solve the problems of low immune protection period of only 7 days, poor stability of attenuated strains, and large number of passages , to achieve the effect of saving labor costs, high commercial value, and less stress on the fish body

Inactive Publication Date: 2015-09-16
GUANGXI ACADEMY OF FISHERY SCI
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  • Abstract
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AI Technical Summary

Problems solved by technology

Two methods have been reported to prepare the attenuated Streptococcus agalactiae vaccine: 1) Pridgeon screened out the attenuated Streptococcus agalactiae strain by continuously increasing the concentration of sparfloxain. The virulence is more likely to be strengthened under the stress of no antibiotics, and injection is used for immunization (Pridgeon et al, 2013); 2) Huang used live attenuated Salmonella typhimurium as a vector to express the DNA of Streptococcus agalactiae surface antigen (Sip) Attenuated vaccine, the attenuated strain prepared by this method has poor stability, low immune response intensity, and the immune protection period is only 7 days (Huang et al, 2014)
Continuous passage in vitro is a classic method of attenuating bacterial virulence. The stability and immunogenicity of the obtained attenuated strains are significantly better than those obtained by gene knockout or antibiotic stress screening. The disadvantage is that the number of passages is often large. so time consuming
So far, there are no reports about natural passage of attenuated strains of Streptococcus agalactiae in fish or other hosts

Method used

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  • Tilapia mossambica streptococcus agalactiae oral attenuated freeze-dried vaccine
  • Tilapia mossambica streptococcus agalactiae oral attenuated freeze-dried vaccine
  • Tilapia mossambica streptococcus agalactiae oral attenuated freeze-dried vaccine

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Embodiment Construction

[0030] (1) Preparation of the vaccine:

[0031] The attenuated strain YM001 production seeds stored at -80°C were inoculated on 5% sheep blood agar plate, cultured at 28°C for 36 h, picked a single colony and inoculated in 100 ml optimized improved Martin medium, and cultured at 28°C, 150 rpm for 24 h; All the 100 bacterial cultures obtained from the culture were inoculated into 5000 ml optimized modified Martin medium, and cultured at 28 °C, 150 rpm for 12 h; the cultured bacterial liquid was centrifuged at 9000 g for 5 min, and 350 ml of phosphate buffered saline (PBS) and 50 ml of lyophilization protection were added. Mix well; distribute in 100 10 ml vials, 4 ml / bottle. In addition, use a spectrophotometer and a plate count to count the cultured bacterial solution, and adjust the bacterial solution to make the amount of bacteria in each bottle 5.0 × 10 10 CFU. Then put the subpackaged vials in a freeze dryer for freeze-drying according to the optimized freeze-drying cu...

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Abstract

The invention discloses a tilapia mossambica streptococcus agalactiae oral attenuated freeze-dried vaccine and its preparation method. According to the invention, tilapia mossambica streptococcus agalactiae attenuated bacterial strain YM001 is performed with steps of enlarge cultivation, thalline collection and freeze-drying preparation to obtain the oral attenuated vaccine. The immunizing dose, immunization program and a vaccine usage method of the vaccine can be determined through tests. The immune efficacy, security and maneuverability of the vaccine can be detected and confirmed through field tests, the result shows that the vaccine has the advantages of high protection effectiveness, good security, convenient usage and low cost, the vaccine can be used for preventing tilapia mossambica streptococcus agalactiae, and has high business value.

Description

technical field [0001] The invention relates to a vaccine for preventing tilapia agalactiae streptococcosis, in particular to the bacterial strain used in the vaccine, the preparation of the vaccine, the immunization dosage, the immunization procedure and the immunization method. Background technique [0002] Streptococcus agalactiae is a common pathogenic bacteria in aquaculture, which can harm different species of fish, such as tilapia, sea bass, pomfret, sea bream, grouper, etc. Among them, tilapia streptococcus agalactiae is the most common Severe (Duremdez et al, 2004; Jafar et al, 2008; Amal et al, 2012). The disease causes extremely high mortality and causes huge economic losses to the aquaculture industry every year (Wang et al, 2014). China is the largest tilapia country in the world, and its output accounts for about 45% of the world's total output. For six consecutive years from 2009 to 2014, streptococcal disease broke out in a large area of ​​tilapia aquacultu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/09A61K9/19A23K1/18A23K1/16A61P31/04
Inventor 陈明李莉萍王瑞甘西梁万文唐佳有黄婷雷爱莹罗永巨李健陈福艳
Owner GUANGXI ACADEMY OF FISHERY SCI
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