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An Equine Streptococcus zooepidemicus strain xjmsy16-1 and its application in equine streptococcal disease vaccine

A Streptococcus zooepidemicus and vaccine technology is applied to a equine-origin Streptococcus zooepidemicus strain XJMSY16-1 and its application in the equine Streptococcus disease vaccine, which can solve the needs of the rapid development of the equine industry and is difficult to achieve. Effective control, differences in mutual immunity, etc., to achieve the effect of high added value of products, high market demand, and no danger of scattered poison

Active Publication Date: 2021-02-26
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 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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  • An Equine Streptococcus zooepidemicus strain xjmsy16-1 and its application in equine streptococcal disease vaccine
  • An Equine Streptococcus zooepidemicus strain xjmsy16-1 and its application in equine streptococcal disease vaccine
  • An Equine Streptococcus zooepidemicus strain xjmsy16-1 and its application in equine streptococcal disease vaccine

Examples

Experimental program
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Effect test

Embodiment 1

[0039] Example 1: Culture and biological characteristics of Streptococcus equi subsp. zooepidemicus strain XJMSY16-1 CGMCC No.12428.

[0040] Streptococcus equi subspecies zooepidemicus XJMSY16-1 CGMCC No.12428 is isolated from the pus of streptococcal equi disease horses, 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 0.3-1.2mm, and the colony is round, with smooth surface, neat edges, 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 , d...

Embodiment 2

[0042]Embodiment 2: PCR amplification and sequence determination of the 16S rRNA gene of Streptococcus equi subspecies zooepidemicus strain XJMSY16-1 CGMCC No.12428

[0043] Streptococcus equi subsp. zooepidemic 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 (50mM Tris, 50mM EDTA-Na 2 ) solution for 2 times; mix the cells with 0.5mL TES solution, add an appropriate amount of lysozyme, and incubate at 37°C for 2 hours; add 0.2mL 20% SDS, and incubate at 60°C for 10 minutes; add 0.3mL 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), draw the supernatant, and then use phenol- Treat once with chloroform-isoamyl alcohol (25:24:1); then treat with chloroform-isoamyl alcohol (24:1, v / v) twice until no protein film appears; ...

Embodiment 3

[0048] Embodiment 3: Preparation of inactivated antigen

[0049] 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 present invention relates to a strain of equine Streptococcus zooepidemicus (also known as Streptococcus equi zooepidemic subspecies, Streptococcus equi subsp. zooepidemicus ) XJMSY16‑1, CGMCC No.12428, has the 16S rRNA gene sequence in Sequence Table 1; the strain is isolated from a horse with equine streptococcosis, a Gram-positive bacterium, and the cells are spherical, arranged in pairs, or curved Long chain, poor growth in common broth and plates; β-type hemolysis occurs on 5% sheep blood plates, and a completely transparent hemolysis zone is formed around dewdrop-shaped small colonies, with a width of up to 2.5-3.2 mm. The growth is facultative anaerobic, the growth temperature is from 26°C to 40°C, the optimum growth temperature is 37°C, and the optimum growth pH is 7.4 to 7.5. Ferment glucose to produce acid, but cannot ferment lactose, mannitol and sorbitol. The bacterial strain can be used to prepare inactivated equine streptococcal vaccine. The vaccine prepared by the bacterial strain has good pertinence, low cost, safety and good protection effect.

Description

technical field [0001] The present invention provides a Streptococcus equi subsp. zooepidemicus (Streptococcus equi subsp.zooepidemicus) XJMSY16-1 strain CGMCC No.12428 isolated from the Yili area of ​​Xinjiang, and the use of the strain in the prevention and treatment of streptococcal equine disease Methods and possible applications for the preparation of inactivated vaccines. [0002] technical background [0003] 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. [0004] Streptococcus equi subspecies zooepidemicus streptococcal disease in equine animals is an acute, febrile c...

Claims

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Application Information

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Patent Type & Authority Patents(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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