Staphylococcus aureus and cow mastitis vaccine obtained by inactivating it

A technology for cow mastitis and staphylococcus, applied in the direction of bacteria, antibacterial drugs, bacterial antigen components, etc., can solve the problems of high actual loss, high attack rate of dairy cows, spread of pathogenic microorganisms, etc., and achieve the effect of various dosage forms

Active Publication Date: 2015-10-28
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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  • Staphylococcus aureus and cow mastitis vaccine obtained by inactivating it
  • Staphylococcus aureus and cow mastitis vaccine obtained by inactivating it

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The acquisition and preservation of embodiment 1, bacterial strain

[0035] 1. Screening of strains

[0036] 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.

[0037] Isolation and purification of Staphylococcus aureus strains from milk samples. Colony morphological characteristics of Staphylococcus aureus (37°C, cultured ...

Embodiment 2

[0051] Embodiment 2, the characteristic of bacterial strain

[0052] The cryopreservation tube obtained in Step 2 of Example 1 and 3 was thawed at room temperature, and the P3 generation strain was subjected to the following steps:

[0053] 1. Immunogenicity of strains

[0054] Rabbits were immunized with the P3 generation strain SACP5-9 (single immunization, each by subcutaneous injection of 5×10 7CFU / 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 SACP5-9 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 negativ...

Embodiment 3

[0072] Embodiment 3, the preparation of vaccine

[0073] 1. Preparation of capsular polysaccharide

[0074] The cryopreservation tube obtained in Step 2 of Example 1 and 3 was thawed at room temperature, and the P3 generation strain was subjected to the following steps:

[0075] 1. Take 3-5 typical colonies of strain SACP5-9, inoculate them in BHI medium, and cultivate them with shaking at 37°C and 150rpm for 18h.

[0076] 2. Take 2 parts by volume of the bacterial liquid obtained in step 1, inoculate 100 parts by volume of BHI medium, and ferment in a fermenter at 37°C (at the initial stage of fermentation, the bacterial concentration of strain SACP5-9 is 2×10 8 per ml, the temperature of the fermenter is 37°C, the initial rotation speed of the fermenter is 198r / min, the tank pressure is maintained at 0.05MPa, and the pH of the fermentation system is adjusted to 7.0-7.2 with 8% NaOH aqueous solution or 5% hydrochloric acid aqueous solution. Rotate the speed of the fermenter...

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Abstract

The invention discloses Staphylococcus aureus strains and a milk cattle mastitis vaccine comprising inactivated Staphylococcus aureus strains. The milk cattle mastitis vaccine comprises inactivated strain SACP5-9, inactivated SACP8-6 and inactivated SACP336-3 as active ingredients. The milk cattle mastitis vaccine comprises inactivated strains and capsular polysaccharides as active ingredients, wherein the inactivated strains include inactivated strain SACP5-9, inactivated SACP8-6 and inactivated SACP336-3; capsular polysaccharides include capsular polysaccharide of strain SACP5-9, capsular polysaccharide of SACP8-6 and capsular polysaccharide of SACP336-3. The vaccine provided by the invention has the advantages of high efficiency, general use, good safety and stability, a variety of dosage forms, and the like. The milk cattle mastitis vaccine provides strong guarantee for safely, effectively and comprehensively preventing Staphylococcus aureus injection type milk cattle mastitis, and is a new milestone of development of Staphylococcus aureus mastitis vaccines.

Description

technical field [0001] The invention relates to staphylococcus aureus and a cow mastitis vaccine obtained by inactivating the staphylococcus aureus. Background technique [0002] 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. The morbidity and mortality of dairy cow mastitis caused by infection are 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 type are: fever, swelling a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/085A61K39/116A61P31/04A61P15/14C12N1/20C12R1/445
Inventor 李敏王林叶孙治华王欢杨鹏辉段跃强孙科孙艺萍
Owner INNER MONGOLIA HUAXI BIOTECH
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