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Mycoplasma hyopneumoniae attenuated strain and application thereof in preparation of attenuated vaccines

A technology for mycoplasma hyopneumoniae and attenuated vaccines, which is applied to attenuated strains of mycoplasma hyopneumoniae and its application in the preparation of attenuated vaccines, and can solve the problems of unsatisfactory protective effects of inactivated vaccines, undetectable serum antibodies, and difficulty in clinical application. , to achieve good protective effect, good immune effect and strong immune protection

Active Publication Date: 2020-12-29
HUAZHONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

More and more studies have proved that the protective effect of inactivated vaccines is not very satisfactory, and the corresponding serum antibodies cannot be detected after immunization with inactivated vaccines, because the immune mechanism induced by Mycoplasma hyopneumoniae is mainly cellular immunity and mucosal immunity
However, the attenuated vaccine commonly used in China requires intrapulmonary injection, which is complicated to operate and not easy for clinical application; moreover, the susceptibility of local breeding pigs to mycoplasma is much higher than that of commercial pigs, and the ideal strain selection should be local isolates

Method used

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  • Mycoplasma hyopneumoniae attenuated strain and application thereof in preparation of attenuated vaccines
  • Mycoplasma hyopneumoniae attenuated strain and application thereof in preparation of attenuated vaccines
  • Mycoplasma hyopneumoniae attenuated strain and application thereof in preparation of attenuated vaccines

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Effect test

Embodiment 1

[0024] The Mycoplasma hyopneumoniae ES-2L strain of the present invention is based on the local clinical isolation of the highly virulent strain ES-2 strain (preservation number CCTCC NO: M 2018570), which is continuously passed in vitro for 200 generations. The specific subculture process is as follows: resuscitate and activate the frozen ES-2 strain, and use the resuscitated and activated bacterial liquid as the P1 generation; After 2 to 3 days of cultivation in the environment, harvest it as the P2 generation; carry out continuous subculture in this way, and when it reaches the 197th generation, spread the harvested bacterial liquid on the improved Friis solid medium, and place it in an environment of 37°C and 5% carbon dioxide After 7-10 days of medium culture, pick a single colony under a low-magnification microscope and insert it into Friis liquid medium for culture; carry out 3 consecutive clone purification and identification in this way, and name the bacterial liquid o...

Embodiment 2

[0036] The P1 generation and ES-2L strain of Mycoplasma hyopneumoniae ES-2 strain were evaluated for virulence. Twelve 25-day-old Landrace male pigs and Large White sow binary hybrid pigs were divided into three groups, A, B, and C, with 4 pigs in each group. Among them, the P1 generation Mycoplasma hyopneumoniae of the P1 generation was injected intratracheally in the A group. Group B was injected with intratracheal injection of Mycoplasma hyopneumoniae ES-2L strain, and group C was injected with normal saline (blank control group). The attacking doses of group A and group B were both 7×10 8 CCU. After the challenge, the animals were dissected after 42 days of observation, during which the body temperature, body weight and clinical symptoms were observed.

[0037] The virulence evaluation experiment showed that there was no increase in body temperature in each group after the challenge; the body weight was measured and the daily average gain was calculated. -2L strain chal...

Embodiment 3

[0043] The Mycoplasma hyopneumoniae ES-2L strain was cultivated to a stable phase in an environment of 37°C and 5% carbon dioxide with the improved Friis liquid medium, and the bacterial liquid was harvested, and the harvested bacterial liquid was diluted with the improved Friis liquid medium to adjust the titer to 1 ×10 7 CCU / mL, prepared into attenuated vaccine. Fifteen 14-day-old Landrace boars and Large White sows were divided into groups A, B, and C, with 5 pigs in each group. Group A is the intramuscular injection immunization group, 14 days old neck intramuscular injection immunization once, 14 days after the first immunization, booster immunization once with the same dose and procedure, each time 2mL / head; B group is the normal saline immunization group (challenge virus) control group), and group C was the blank control group. Immunization dose is 2×10 7 In CCU, except for the blank control group, the other two groups received intratracheal injection of 7 mL of viru...

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Abstract

The invention discloses a mycoplasma hyopneumoniae attenuated strain and application thereof in preparation of attenuated vaccines, and belongs to the technical field of microorganisms and immunity. The mycoplasma hyopneumoniae attenuated strain is classified and named as mycoplasma hyopneumoniae ES-2L, the preservation number is CCTCC NO: M 2020313, and the mycoplasma hyopneumoniae attenuated strain has the characteristics of high growth speed, high growth titer, good safety, strong immune protection capability, direct intramuscular injection immunization and the like, and can be used for preparing the mycoplasma hyopneumoniae attenuated vaccines. The mycoplasma hyopneumoniae attenuated vaccine prepared based on the vaccine strain has the advantages of being convenient to use clinically,high in safety, good in immune effect and the like.

Description

technical field [0001] The invention belongs to the technical field of microorganisms and immunization, and in particular relates to an attenuated strain of Mycoplasma hyopneumoniae and its application in preparing an attenuated vaccine. Background technique [0002] Mycoplasma hyopneumoniae is a respiratory pathogen that can cause porcine enzootic pneumonia. The disease is also known as "swine asthma". It is a chronic respiratory disease that mainly infects pigs. Pigs of all ages can be infected. Clinical symptoms Manifested as coughing and wheezing, emaciation, histological lesions mainly manifested as catarrhal exudate in the trachea and substantial "meat change" in the lung tissue, once the lesion appeared, it would be difficult to recover. In the lungs, especially the apical lobe, heart lobe, and middle lobe, there are light red and gray red "meat changes". As the course of the disease progresses, off-white, light red "shrimp-like consolidation" will appear. The disea...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K39/02A61P31/04C12R1/35
CPCA61K39/0241A61P31/04A61K2039/522A61K2039/70C12R2001/35C12N1/205
Inventor 谭臣李真亚陈焕春
Owner HUAZHONG AGRICULTURAL UNIVERSITY
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