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A Streptococcus equine epidemic strain xz1 and its application in equine epidemic vaccine

A technology of Streptococcus equine and equine adenovirus, applied in the direction of bacteria, antibacterial drugs, bacterial antigen components, etc., can solve the needs of the rapid development of the horse industry, the difference in mutual immunity, and it is difficult to achieve effective control, etc. problems, to achieve the effect of high added value of products, no danger of scattered poison, and high market demand

Active Publication Date: 2016-02-24
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 Streptococcus equine epidemic vaccines in China, which cannot meet the needs of the rapid development of the equine industry.
In addition, because the strains prevailing in different regions are not the same, the mutual immunity between different epidemic strains will also have certain differences. In addition, it is also found that such strains have the characteristics of variation, and it is difficult to effectively control the strains in different regions with only one vaccine strain. occurrence of disease

Method used

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  • A Streptococcus equine epidemic strain xz1 and its application in equine epidemic vaccine
  • A Streptococcus equine epidemic strain xz1 and its application in equine epidemic vaccine
  • A Streptococcus equine epidemic strain xz1 and its application in equine epidemic vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Culture and biological characteristics of Streptococcus equine epidemic strain XZ1CGMCC No.7830.

[0037] Streptococcus equi subspecies XZ1CGMCCNo.7830 is isolated from the pus of horses with equine blight disease, inoculated in blood as the substrate medium, and cultured at 37°C for 12-15h. The bacterium has the following characteristics: (1) Colony characteristics: the diameter of the colony cultivated on the blood agar plate for 1 day is 0.2-1.0mm, and the colony is round, with smooth surface, neat edges, raised, off-white. (2) Morphological characteristics of cells: cells are round or oval; Gram staining is positive; cell diameter is 1.0-2.0 μm. (3) Physiological and biochemical characteristics: facultative anaerobic growth; growth temperature ranges from 28°C to 39°C, optimum growth temperature is 37°C; pH growth range is 6-8, optimum growth pH is 7.4-7.6; can ferment glucose, sucrose , does not hydrolyze aescin, does not grow in 6.5% NaC1, does not liq...

Embodiment 2

[0039] Example 2: PCR amplification and sequence determination of the 16SrRNA gene of Streptococcus equine epidemic strain XZ1CGMCCNo.7830

[0040] Streptococcus equine epidemic strain XZ1CGMCCNo.7830 was inoculated in liquid medium, and the fermentation broth grown to the late logarithmic period was centrifuged (5000 rpm, 5 minutes) to remove the supernatant, and TE (50mM Tris, 50mM EDTA-Na 2 ) solution for 2 times; mix the cells with 0.5mLTES solution, add appropriate amount of lysozyme, and keep warm at 37°C for 2 hours; add 0.2mL 20% SDS, keep warm at 60°C for 10 minutes; add 0.3mL 5MNaClO 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 was added to 20μl 0.2% RNase was i...

Embodiment 3

[0045] Embodiment 3: Preparation of inactivated antigen

[0046] The isolated bacteria were respectively inoculated in TM medium supplemented with 2% horse serum, cultured at 37°C and 180r / min for 18-24 hours, and inactivated by adding 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 sterile PBS (pH 7.2, 10 mmol) 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 a strain of strangles streptococcus (also known as streptococcus. equi subsp equi) XZ1 with the preservation number of CGMCC No.7830. The strangles streptococcus strain has the16S rRNA gene sequence shown in the sequence table 1; the strain is isolated from sick horses with strangles and has gram positive bacteria, and cells are spherical and are arranged in pairs or cells are bent chain-shaped, which are in poor growth in ordinary broth and plate; beta type hemolysis happens on a plate with 5% of sheep blood, and a completely transparent hemolytic tape with the width of 3 mm can be formed around small dewdrop-shaped bacterial colonies; facultative anaerobic growth can be realized with the growth temperature from 25 DEG C to 40 DEG C, and the optimum growth temperature is 37 DEG C, and the optimum pH value for growth is between 7.4 and 7.5; glucose is fermented for production of acid, and lactose, mannitol and sorbitol cannot be fermented. The strangles streptococcus strain can be used for preparation of strangles inactivated vaccine, and the prepared strangles inactivated vaccine is good in pertinence, low in cost, safe and good in protection effect.

Description

technical field [0001] The present invention provides a Streptococcus equisubspequi XZ1 strain CGMCC No.7830 isolated from Ili region of Xinjiang, and the method and possible application of the strain for preparing inactivated vaccine in the prevention and treatment of equine epidemic. technical background [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. The rapid development of the horse industry and the increasing scale and intensification of horse breeding have made the hazards of horse gland disease more and more obvious. [0003] Stangles is an acute contagious disease of equine animals caused by Streptococcus equi, mainly in the neck and head of horses with suppurative lymphangitis. The disease is commonly known as jet throat or groove knot, lar...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K39/09A61P31/04C12R1/46
Inventor 苏艳王彩蝶刘云涛张宝江
Owner XINJIANG AGRI UNIV
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