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A Salmonella abortus equine strain smxj-97 and its application in equine abortus salmonella vaccine

A technology of SMXJ-97 and Salmonella, which is applied in the field of preparation of inactivated vaccines, can solve the problems of not being able to meet the needs of the rapid development of the horse industry, differences in mutual immunity, and difficulty in achieving effective control, and achieve high product added value and no scattered vaccines. Toxic danger, high market demand effect

Active Publication Date: 2021-03-02
XINJIANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, there are very few researches and products on the commercialization of vaccines for equine abortive salmonellosis in China, which are far from meeting 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. It is difficult to effectively control the occurrence of the disease in different regions with only one vaccine strain.

Method used

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  • A Salmonella abortus equine strain smxj-97 and its application in equine abortus salmonella vaccine
  • A Salmonella abortus equine strain smxj-97 and its application in equine abortus salmonella vaccine
  • A Salmonella abortus equine strain smxj-97 and its application in equine abortus salmonella vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Culture and biological characteristics of Salmonella equine abortus strain SMXJ-97CGMCC No.9047.

[0037]The Salmonella abortus equine strain SMXJ-97CGMCC No.9047 was isolated from the fetus of an aborted horse, inoculated in MM medium, and cultured at 37°C for 12-15h. The bacterium has the following characteristics: (1) Colony characteristics: white colonies with round shape, smooth surface, protrusions, neat edges, and uniform shape and size grow on MM medium. (2) Morphological characteristics of the cells: the bacteria are straight rods with a size of 0.7-1.5 μm×2.0-5 μm, and Gram staining is negative. (3) Physiological and biochemical characteristics: aerobic and facultative anaerobic bacteria can grow on ordinary culture medium, the growth temperature is from 25°C to 40°C, the optimum temperature is 37°C, the pH growth range is 5-9, the optimum PH7.4~7.6. Can ferment mannitol, decompose lysine, do not decompose urea, cannot use tryptophan, malonate, sa...

Embodiment 2

[0039] Embodiment 2: PCR amplification and sequence determination of the 16S rRNA gene of Salmonella equine abortus bacterial strain SMXJ-97CGMCC No.9047

[0040] Salmonella equine abortus strain SMXJ-97CGMCC No.9047 was inoculated in liquid medium, and the fermentation broth grown to the late logarithmic period was centrifuged (5000rpm / min, 5min) 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 2h; add 0.2mL 20% SDS, and incubate at 60°C for 10min; 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; add 20 μL 0.2% RNA to the supernatant T...

Embodiment 3

[0045] Embodiment 3: Preparation of inactivated antigen

[0046] The isolated bacteria were respectively inoculated in MM 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 Salmonella abortus equine ( Salmonella abortus equi ) SMXJ‑97 CGMCC No.9047, which has the 16S rRNA gene sequence in Sequence Table 1; the strain was isolated from aborted horses in Xinjiang, and the bacteria were straight rod-shaped, with a size of 0.7~1.6μm×2.0~5μm, Gram Stain negative. It is an aerobic and facultative anaerobic bacterium, which can grow on ordinary culture medium. The growth temperature is from 25°C to 40°C, the optimum temperature is 37°C, the pH growth range is 5-9, and the optimum pH is 7.4-7.6. It can ferment mannitol, decompose lysine, but not urea, and cannot use tryptophan, malonate, salicin, and sorbitol. The strain can be used to prepare an inactivated vaccine against salmonellosis in equine abortion. 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 strain of Salmonella abortusequi (Salmonella abortusequi) SMXJ-97 strain CGMCC No.9047 isolated from the Yili area of ​​Xinjiang, and the method and possible application of the strain for preparing inactivated vaccines in the prevention and treatment of salmonellosis in equine abortusequi . [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 abortive salmonellosis in horses are becoming more and more obvious. [0004] Salmonella abortus equine, also known as equine paratyphoid, is an infectious disease of equine animals caused by Salmonella abor...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K39/112A61P31/04C12R1/42
CPCA61K39/0275A61K2039/521C12N1/205C12R2001/42
Inventor 苏艳杨康苏玲玲
Owner XINJIANG AGRI UNIV
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