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A Porcine Pathogenic Escherichia coli Strain and Its Inactivated Vaccine

A technology of Escherichia coli and inactivated vaccine, applied in vaccines, veterinary vaccines, bacteria and other directions to achieve good immune protection effect and good protection effect

Active Publication Date: 2020-12-15
HENGYANG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far there is no commercially available E. coli vaccine or related drugs that can prevent multiple serotypes of ETEC in China.

Method used

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  • A Porcine Pathogenic Escherichia coli Strain and Its Inactivated Vaccine
  • A Porcine Pathogenic Escherichia coli Strain and Its Inactivated Vaccine
  • A Porcine Pathogenic Escherichia coli Strain and Its Inactivated Vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Isolation, Identification and Preservation of Porcine Pathogenic Escherichia coli

[0029] 1.1 Collection of disease materials

[0030] In July 2016, in a medium-sized farm on the outskirts of Hengyang City, severe diarrhea occurred in multiple pens of piglets within two months, and the disease could not be controlled after antibiotic treatment, and the mortality rate was as high as 20%. Aseptically collect the intestinal tissue and intestinal contents of dead piglets, streak the samples on the MAC plate, place them upside down in a constant temperature incubator at 37°C for 12-16 hours, and then pick a single colony of suspected E. coli in pink or red color for inoculation On eosin melanin medium, cultivate in a constant temperature incubator at 37°C for 12-16h, pick a single purple-black colony with metallic luster and inoculate it in an LB test tube, culture on a shaker at 160r / min at 37°C for 16h, mix with 50% glycerol 1 : 1 mixed and stored at -40°C for...

Embodiment 2

[0040] Embodiment 2: the preparation of porcine pathogenic escherichia coli inactivated vaccine

[0041] 2.1 Selection of optimal bacterial content in inactivated vaccines

[0042] Take 40 piglets aged 4 weeks and divide them into 5 groups with 8 piglets in each group. Among them, the vaccine immunization test group 1-4 and the control test group, the immunization dose of the immunization test group is 1×10 9 CFU / portion, 5×10 8 CFU / portion, 2.5×10 8 CFU / portion, 1.25×10 8 CFU / head (for the preparation method of the vaccine, please refer to the preparation method of inactivated vaccine in 2.2 below), the control group was injected with the same dose of PBS, and the same feeding management conditions were adopted, and a booster immunization was given 14 days after the first immunization. On the 10th day after each immunization, the blood was collected separately, and the serum was separated, and the antibody level in the serum was detected by indirect ELISA method, and the f...

Embodiment 3

[0055] Example 3: Immunological efficacy evaluation of porcine pathogenic Escherichia coli inactivated vaccine

[0056] 3.1 Comparison of immune efficacy of inactivated vaccines

[0057] 3.1.1 Test materials: E.coli-HuNHY19 (CGMCC No.17622), E.coli-HuNHY18 (self-isolated), Escherichia coli CVCC1524 (O8 serotype, purchased from China Veterinary Microorganism Strain Collection Center CVCC).

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Abstract

The present invention isolates a highly pathogenic and highly lethal porcine pathogenic Escherichia coli E.coli-HuNHY19 from a pig farm with severe disease, and inactivates it to prepare an inactivated vaccine, which has the effect on piglets Good immune protection effect, which can realize the complete protection of piglets against the E. coli, also has a good protective effect on the strain of the same serotype O8, and has a certain immune cross-protection effect on other serotypes. Through sows Immunization experiments found that the inactivated vaccine can well stimulate primiparous sows to produce antibodies, and the antibody level and attenuation speed in their colostrum are extremely superior to the existing isolated strains of the same serotype, and can achieve the effect on lactation. Piglets are better protected.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a porcine pathogenic Escherichia coli strain and an inactivated vaccine thereof. Background technique [0002] Escherichia coli is a component of the symbiotic flora in the intestinal tract of humans and animals, and some pathogenic Escherichia coli can cause various symptoms such as diarrhea and sepsis in humans and animals. Porcine colibacillosis is an intestinal infectious disease of piglets caused by pathogenic Escherichia coli. Among them, yellow scour of piglets, pullorum of piglets and piglet edema are common clinical symptoms, and it is caused by other causes of Escherichia coli. Diarrhea etc. Yellow dysentery of piglets is mainly manifested as pulling yellow or yellow-green thin feces, which contain curd flakes and small bubbles, and are foul-smelling. Piglets lose weight rapidly after diarrhea, dehydrate and die, and the mortality rate can be as high as more t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K39/108A61P31/04A61P1/12C12R1/19
CPCA61K39/0258A61K2039/521A61K2039/552A61P1/12A61P31/04C12N1/205C12R2001/19
Inventor 唐青海滕威杨海阳坤王芳宇杨灿何丽芳唐娇玉曹丽敏易诚唐斯萍
Owner HENGYANG NORMAL UNIV