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Serotype 7 haemophilus parasuis natural weak virulent strain and application thereof

A Haemophilus suis, natural technology, used in bacteria, antibacterial drugs, medical preparations containing active ingredients, etc. The effect of high drug production efficiency, simple preparation method and weakened pathogenicity

Active Publication Date: 2018-05-11
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there have been instances of prophylaxis failure with inactivated vaccines, possibly due to lack of cross-protection due to different serotypes of the causative and vaccine strains
While cross-protection is an essential requirement for commercial inactivated vaccines, strain-specific inactivated vaccines may lack potency due to the presence of more than one strain or serotype in the herd, or due to the subsequent introduction of new strains into the herd lose effectiveness
Due to the diversity of Haemophilus parasuis serotypes and a large proportion of strains that cannot be typed, the differences in virulence of the strains, and the current lack of deep understanding of protective antigens and virulence factors, it is not possible to have an inactivated strain Seedlings are simultaneously cross-protective against all pathogenic strains

Method used

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  • Serotype 7 haemophilus parasuis natural weak virulent strain and application thereof
  • Serotype 7 haemophilus parasuis natural weak virulent strain and application thereof
  • Serotype 7 haemophilus parasuis natural weak virulent strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Isolation and identification of natural attenuated strain of Haemophilus parasuis type 7:

[0029] 1) Strains

[0030] Haemophilus parasuis type 7 natural attenuated strain, isolation number HPS1712, was isolated from the nasal cavity of a healthy piglet in a pig farm in Wuyang County, Henan Province on February 6, 2007, and was isolated and identified by the State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University and save.

[0031] Haemophilus parasuis type 5 SH0165 strain, type 4 MD0322 strain, and type 13 FX2002 strain were isolated, identified and preserved by the State Key Laboratory of Agricultural Microbiology of Huazhong Agricultural University as the strains for testing.

[0032] 2) Experimental animals

[0033] 16-20g healthy Balb / c mice; 28-35-day-old healthy piglets, purchased from Hubei Wuhan Wannianqing Animal Husbandry Co., Ltd., tested by Haemophilus parasuis antibody ELISA detection kit (self-built method) and the antibody ...

Embodiment 2

[0058] The Haemophilus parasuis type 7 natural attenuated strain HPS1712 that embodiment 1 obtains carries out pathogenicity detection:

[0059] 1) Determination of toxicity to mice

[0060] 130 healthy Balb / c mice weighing 16-20 g were randomly divided into 13 groups, 10 in each group. The Haemophilus parasuis HPS1712 strain and the virulent strain SH0165 were divided into 6 gradients according to different doses, each strain was injected into 6 groups of mice, and the bacterial solution was injected into the chest cavity with 0.5ml / mouse, and the number of viable bacteria was 7.7 ×10 7 CFU, 9.4×10 8 CFU, 1.3×10 9 CFU, 1.7×10 10 CFU, 2.1×10 11 CFU, 1.7×10 12 CFU, another group was injected with 0.5ml PBS as a control, observed for 14 days after the injection, and counted the mortality of mice in each group, the results are shown in Table 4 below.

[0061] Table 4 Haemophilus parasuis type 7 HPS1712 strain and virulent strain SH0165 strain are to the infection test resu...

Embodiment 3

[0069] Detection of immune efficacy of Haemophilus parasuis type 7 natural attenuated strain HPS1712:

[0070] 1) Preparation of the vaccine:

[0071] S1. Cell culture:

[0072] Inoculate the bacterial powder of Haemophilus parasuis HPS1712 strain on the TSA / V / S solid medium, and incubate at 37°C for 24 hours; pick a single colony and re-inoculate the TSA / V / S solid medium for 3 to 5 generations. Incubate at 37°C for 12-16 hours, then pick a single colony on the plate and inoculate it in TSB / V / S liquid medium for 12 hours of activation. After passing the pure inspection, transfer the single colony according to the volume ratio of the bacterial solution to the medium 1:100 To a large bottle of TSB / V / S liquid medium, 37 ° C, 170rpm shaker shaker expansion culture for 12h, the TSA / V / S and TSB / V / S medium is a medium containing 5% bovine serum, 0.2% NAD ;

[0073] S2. Cell collection:

[0074] Transfer the bacterial solution cultured in the liquid in step S1 into a 50ml sterile ...

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Abstract

The invention discloses a serotype 7 haemophilus parasuis natural weak virulent strain and application thereof. The serotype 7 haemophilus parasuis natural weak virulent strain is deposited at China Center for Type Culture Collection on November 22, 2017, and the accession number is CCTCC M 2017706. The serotype 7 haemophilus parasuis natural weak virulent strain has very low pathogenicity and issuitable for preparation of a medicament for treatment of haemophilus parasuis, and the medicament is a vaccine. A preparation method of the vaccine includes the following steps: S1, thallus culture;S2, thallus collection; S3, freeze-drying. Inactivation and inactivation inspection are not needed in the preparation of the medicament, the preparation method is simple, and production efficiency ofthe medicament is higher. When the vaccine is applied to 28 to 35-day-old piglets in immunizing dose of 2.0*108CFU / mL, the immune protection rate is 100%.

Description

technical field [0001] The invention belongs to the application field of Haemophilus parasuis, more specifically, relates to a natural attenuated strain of Haemophilus parasuis serotype 7 and its application. Background technique [0002] Haemophilus parasuis disease is a new bacterial infectious disease that seriously harms the pig industry in recent years, causing multiple serositis, arthritis and meningoencephalitis in pigs. The incidence rate is generally 10-15%, and the mortality rate It can reach more than 50%, which has brought huge economic losses to the pig industry, and has aroused people's increasing attention. The serotypes of Haemophilus parasuis (Hps), the pathogenic bacterium of the disease, are complex and diverse. According to the Kieleetin. The serotype of more than % of the isolates could not be determined. Existing data show that Hps has obvious endemic characteristics. [0003] Because Haemophilus parasuis is highly susceptible to drug resistance, an ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K39/102A61P31/04C12R1/21
CPCA61K39/102C12N1/205C12R2001/21
Inventor 王湘如蔡旭旺张勤学陈焕春徐晓娟
Owner HUAZHONG AGRI UNIV
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