A strain of Escherichia coli and a cow mastitis vaccine obtained by inactivating the strain

A technology for dairy cow mastitis and Escherichia coli, which is applied in the direction of bacteria, antibacterial drugs, and methods based on microorganisms, can solve the problems of high actual loss, spread of pathogenic microorganisms, and high attack rate of dairy cows, and achieve the effect of various dosage forms

Active Publication Date: 2016-06-22
INNER MONGOLIA HUAXI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The recessive type has no specific clinical manifestations and can only be detected through somatic cells, milk production and quality testing, but the attack rate of dairy cows is much higher than that of the clinical type, causing further spread of pathogenic microorganisms, and under certain conditions it will transform into clinical symptoms. type, the actual loss is much higher than the clinical type

Method used

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  • A strain of Escherichia coli and a cow mastitis vaccine obtained by inactivating the strain
  • A strain of Escherichia coli and a cow mastitis vaccine obtained by inactivating the strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1, acquisition and preservation of recombinant bacteria

[0030] 1. Screen wild bacteria

[0031] 30,000 cow milk samples were collected from 90 medium and large dairy farms in 30 provinces and cities including Inner Mongolia, Chongqing, Guangzhou, Heilongjiang, Lanzhou, Hebei, and Shandong. Milk sample collection method of dairy cows: select cows with clinical symptoms of mastitis (or suspected subclinical mastitis), first scrub the udders with warm water, then scrub the udders with 0.2% bromogeramine, and finally wipe the nipples with 70% alcohol. At the same time, use 70% alcohol to wipe and disinfect the fingers. Squeeze out the first 2-3 handfuls of milk from each milk chamber to eliminate contaminated bacteria. Take at least 5mL of milk samples from each cow in a sterile milk sample cup.

[0032] Isolation and purification of Escherichia coli strains from milk samples. Colony morphological characteristics of Escherichia coli (37°C, cultured for 24-72h...

Embodiment 2

[0041] Embodiment 2, the characteristic of bacterial strain EBMO9

[0042] The cryopreservation tube obtained in step 2 of Example 1 was thawed at room temperature, and the P3 generation strain EBMO9 was subjected to the following steps:

[0043] 1. Immunogenicity of strain EBMO9

[0044] Rabbits were immunized with the P3 generation strain EBMO9 (single immunization, each by subcutaneous injection of 5×10 7 CFU / ml, 1 ml for each injection), 21 days later, blood was collected from the heart, and the serum was separated. The IgG antibody titer of the serum was detected by ELISA (serum dilution was obtained by gradient dilution with PBST buffer; the P3 generation strain EBMO9 was used as the coating source, and the coating concentration was 1×10 9 Bacteria / ml; goat anti-rabbit IgG enzyme-labeled secondary antibody; set negative control, that is, replace serum diluent with PBST buffer; detect the absorbance value at 450nm; the absorbance value is more than 2.1 times that of the...

Embodiment 3

[0051] Embodiment 3, the preparation of vaccine

[0052] The cryopreservation tube obtained in step 2 of Example 1 was thawed at room temperature, and the P3 generation strain EBMO9 was subjected to the following steps:

[0053] 1. Take the P3 generation strain EBMO9 and resuspend it with PBS buffer.

[0054] 2. Take the bacterial solution obtained in step 1, add 37-40% formaldehyde solution to make the formaldehyde concentration 0.4% (volume ratio), incubate at 37°C and 150rpm for 36h (36h-48h is acceptable in practical applications), centrifuge (5000r / min, 20min) to collect the bacteria, wash the bacteria with sterilized PBS buffer, and then suspend the bacteria with sterilized PBS buffer to obtain Vaccine-I (containing 1×10 10 bacteria / mL).

[0055] 3. Take vaccine-I, add aluminum hydroxide to make the concentration 0.5 mg / ml, and obtain vaccine-II.

[0056] 4. Take vaccine-I, add aluminum phosphate to make the concentration 0.5 mg / ml, and obtain vaccine-III.

[0057] ...

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Abstract

The invention discloses an Escherichia coli strain and a dairy cow mastitis vaccine obtained by inactivating the same. The Escherichia coli (Escherichia coli) EBMO9 provided in the invention is called strain EBMO9 for short and has the collection number of CGMCC No. 8532. The invention also provides application of the strain EBMO9 in preparation of the dairy cow mastitis vaccine. The invention further provides a dairy cow mastitis vaccine which has an active ingredient of the inactivated strain EBMO9. A powerful guarantee is provided for safely, efficiently, comprehensively and effectively preventing Escherichia coli infected dairy cow mastitis, and a novel milestone is constructed for development of Escherichia coli mastitis vaccines.

Description

technical field [0001] The invention relates to an Escherichia coli strain and a cow mastitis vaccine obtained by inactivating the strain. Background technique [0002] Cow mastitis (Bovine Mastitis) is a major infectious disease that endangers the development of animal husbandry and food safety. Especially in recent years, with the change of the environment and the abuse of antibiotics, a variety of drug-resistant bacteria and even superbugs have emerged. Bacterial infections The morbidity and mortality of dairy cow mastitis caused by it have been increasing year by year, seriously endangering the development of animal husbandry and food safety, and further endangering human health. The economic losses caused by cow mastitis amount to 3 billion U.S. dollars in the United States, 400 million U.S. dollars in the United Kingdom, and more than 3.5 billion U.S. dollars in my country. Cow mastitis is divided into clinical type and recessive type. The symptoms of the clinical ty...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K39/108A61P31/04A61P15/14C12R1/19
Inventor 王欢王林叶孙治华孙科李敏孙艺萍段跃强杨鹏辉
Owner INNER MONGOLIA HUAXI BIOTECH
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