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Salmonella abortus equi strain SMXJ-97 and application thereof in salmonella abortus equi 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. Poison hazard, high market demand effect

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

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, since 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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  • Salmonella abortus equi strain SMXJ-97 and application thereof in salmonella abortus equi vaccine
  • Salmonella abortus equi strain SMXJ-97 and application thereof in salmonella abortus equi vaccine

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Example 1: Culture and biological characteristics of Salmonella equine abortus strain SMXJ-97 CGMCC No.9047.

[0039]Salmonella abortus equine strain SMXJ-97 CGMCC No.9047 was isolated from the fetus of aborted horse, inoculated in MM medium, and cultured at 37 ℃ for 12-15 h. The bacteria has the following characteristics: (1) Colony characteristics: On MM medium, it grows into white colonies with round shape, smooth surface, protrusions, neat edges, and uniform shape and size. (2) Cell morphological characteristics: The bacteria are straight rod-shaped, with a size of 0.7-1.5 μm×2.0-5 μm, and Gram staining is negative. (3) Physiological and biochemical characteristics: It is an aerobic and facultative anaerobic bacteria, which can grow on ordinary culture medium, the growth temperature is from 25 ℃ to 40 ℃, the optimum temperature is 37 ℃, the pH growth range is 5-9, the optimum PH7.4~7.6. Can ferment mannitol, decompose lysine, do not decompose urea, cannot use tryp...

Embodiment 2

[0041] Embodiment 2: PCR amplification and sequence determination of the 16S rRNA gene of Salmonella abortus strain SMXJ-97 CGMCC No.9047

[0042] Salmonella equine abortus strain SMXJ-97 CGMCC No.9047 was inoculated in liquid medium, and the fermentation broth grown to the late logarithmic period was centrifuged (5000 rpm / min, 5 min) 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 h; add 0.2 mL 20% SDS, and incubate at 60 ℃ for 10 min; add 0.3 mL 5 M 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 was added with 20 μL 0.2% ...

Embodiment 3

[0047] Embodiment 3: Preparation of inactivated antigen

[0048] The isolates were inoculated in MM medium supplemented with 2% horse serum, cultured at 37 °C and 180 r / min for 18-24 h, and inactivated with 0.4% formaldehyde for 36-48 h. After complete inactivation, centrifuge at 7000 rpm / min for 15 min to remove the supernatant. The bacterial pellet was diluted to 1×10 with sterile PBS (pH7.2, 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 Salmonella abortus equi strain SMXJ-97 with the preservation number of CGMCC No.9047. The strain has 16S rRNA gene sequences in a sequence table 1. The stain is separated from abortion sick horses in Xinjiang, the bacterium takes the shapes of a straight rod, the size is 0.7-1.6 [mu]m*2.0-5 [mu]m, and the bacterium is a negative Gram bacterium. The strain is an aerobic facultative anaerobic bacterium and can grow on a common culture medium, the growth temperature is 25-40 DEG C, the most appropriate temperature is 37 DEG C, the pH growth range is 5-9, and the most appropriate pH value is 7.4-7.6. The strain can be adopted to ferment mannitol and decompose lysine, urea cannot be decomposed with the strain, and tryptophan, malonate, saligenin and sorbitol cannot beutilized with the strain. The strain can be used for preparing salmonella abortus equi inactivation vaccines. The vaccines prepared from the strain are high in disease specificity, low in cost, highin security and good in protection effect.

Description

technical field [0001] A strain provided by the invention is isolated from Salmonella abortus equine in Ili area, Xinjiang ( Salmonella abortus equi ) SMXJ-97 strain CGMCC No.9047, and the method and possible application of the strain for preparing inactivated vaccine in the prevention and treatment of aborted salmonellosis in equine. 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 abortive salmonellosis in horses are becoming more and more obvious. [0003] Salmonella abortus equine, also known as equine paratyphoid, is caused by Salmonella abortus equine ( Salmonella abortus equi ) is an infectiou...

Claims

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Application Information

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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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