Streptococcus equi strain XJMSY16-1 and application thereof in streptococcus equi vaccine

A Streptococcus zooepidemicus and vaccine technology, applied to a strain of Streptococcus zooepidemicus strain XJMSY16-1 from horses and its application in equine streptococcal disease vaccines, can solve the needs of the rapid development of the horse industry, which is difficult to achieve Effective control, mutual immunity differences and other issues, to achieve the effect of high added value of products, high market demand, and no risk of poisoning

Active Publication Date: 2018-06-29
XINJIANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are very few researches and products on the commercialization of equine-sourced Streptococcus zooepidemicus vaccines in China, which cannot meet the needs of the rapid development of the equine industry.
In addition, because the strains prevalent in different regions are not the same, the mutual immunity between different epidemic strains will also have certain differences. In addition, it has been found that such strains have the characteristics of variation, and it is difficult to effectively control the infection in different regions with only one vaccine strain. occurrence of disease

Method used

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  • Streptococcus equi strain XJMSY16-1 and application thereof in streptococcus equi vaccine
  • Streptococcus equi strain XJMSY16-1 and application thereof in streptococcus equi vaccine

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Example 1: Culture and biological characteristics of Streptococcus zooepidemicus strain XJMSY16-1 CGMCC No.12428.

[0039] Streptococcus equi subsp. zooepidemicus XJMSY16-1 CGMCC No.12428 was isolated from the pus of Streptococcus equi diseased horses, inoculated in blood-based medium, and cultured at 37°C for 12-15 hours. The bacteria has the following characteristics: (1) Colony characteristics: the diameter of the colony cultured on the blood agar plate for 1 day is 0.3-1.2 mm, the colony is round, the surface is smooth, the edges are neat, raised, off-white. (2) Morphological characteristics of bacteria: the bacteria are round or oval; Gram staining is positive; the diameter of the bacteria is 1.1-2.1 μm. (3) Physiological and biochemical characteristics: facultative anaerobic growth; growth temperature ranges from 27°C to 39°C, optimum growth temperature is 37°C; pH growth range is 6-8, optimum growth pH is 7.3-7.6; can ferment glucose, sucrose , does not hydrolyz...

Embodiment 2

[0041]Embodiment 2: PCR amplification and sequence determination of the 16S rRNA gene of Streptococcus zooepidemicus strain XJMSY16-1 CGMCC No.12428

[0042] Streptococcus zooepidemicus strain XJMSY16-1 CGMCC No.12428 was inoculated in liquid medium, and the fermentation broth grown to the late logarithmic period was centrifuged (4500 rpm, 5 minutes) to remove the supernatant, and TE (50 mM Tris, 50 mM EDTA-Na 2 ) solution for 2 times; mix the cells with 0.5 mL TES solution, add appropriate amount of lysozyme, and incubate at 37 ℃ for 2 hours; add 0.2 mL 20 % SDS, and incubate at 60 ℃ for 10 minutes; add 0.3 mL 5M NaClO 4 , mix well; add an equal volume of phenol-chloroform-isoamyl alcohol (25: 24: 1), shake gently for about 5 minutes, centrifuge (5000 rpm, 5 minutes), absorb the supernatant, and then use phenol- Chloroform-isoamyl alcohol (25: 24: 1) was treated once; then chloroform-isoamyl alcohol (24: 1, v / v) was treated twice until no protein film appeared; supernatant w...

Embodiment 3

[0047] Embodiment 3: Preparation of inactivated antigen

[0048] The isolates were inoculated in TM medium supplemented with 2% horse serum, cultured at 37°C and 180r / min for 18-24 hours, and inactivated with 0.4% formaldehyde for 36-48 hours. After complete inactivation, centrifuge at 7000rpm / min for 15min to remove the supernatant. The bacterial pellet was diluted to 1×10 with sterilized PBS (pH7.2, 10mmol) 7 CFU / mL. Immunize rabbits after emulsification with the same amount of complete Freund's adjuvant, emulsify with the same concentration of bacteria and the same amount of incomplete Freund's adjuvant 3 weeks later, sterility test and safety test are carried out after the vaccine is fully shaken and mixed.

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Abstract

The invention relates to Streptococcus equi (also named as Streptococcus equi subsp.zooepidemicus) XJMSY16-1 with the preservation number of CGMCC No.12428. The strain has 16S rRNA gene sequences in asequence table 1. The stain is separated from sick horses suffering from Streptococcus equi and is a positive Gram bacterium, cells of the strain take the shapes of spheres and are arranged in pairs,or take the shapes of bent long chains, and are low in growth property in common meat soup and panels; beta-type hemolysis is caused on a panel with 5% fresh sheep blood, completely transparent hemolysis zones are formed around small dewdrop shaped bacterial colonies, and the width of the hemolysis zones can be up to 2.5-3.2 mm. The strain grows in a facultative anaerobic manner, the growth temperature is 26-40 DEG C, the most appropriate temperature is 37 DEG C, and the most appropriate growth pH value is 7.4-7.5. Glucose can be fermented to generate acids, and lactose, mannitol and sorbitolcannot be fermented with the strain. The strain can be used for preparing streptococcus equi deactivation vaccines. The vaccines prepared from the strain are high in disease specificity, low in cost,high in security and good in protection effect.

Description

technical field [0001] A strain provided by the invention is isolated from Streptococcus zooepidemicus ( Streptococcus equi subsp. zooepidemicus ) XJMSY16-1 strain CGMCC No.12428, and the method and possible application of the strain for preparing inactivated vaccine in the prevention and treatment of equine streptococcosis. Background technique [0002] With economic globalization, the modern horse industry has transformed into a huge industry in the form of horse racing, performance exhibitions, riding entertainment, etc., which has ushered in unprecedented development opportunities for the Chinese horse industry. With the rapid development of the horse industry and the increasing scale and intensification of horse breeding, the hazards of equine streptococcosis are becoming more and more obvious. [0003] Streptococcus zooepidemicus caused by Streptococcus zooepidemicus in equine animals is an acute, febrile contact infectious disease, which mainly manifests as lower r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K39/09A61P31/04C12R1/46
CPCA61K39/092A61K2039/521A61K2039/552C12N1/205C12R2001/46
Inventor 苏艳易鸳鸯苏玲玲
Owner XINJIANG AGRI UNIV
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