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The Research of the of the Virulence Factors of Staphylococcus Aureus in Dairy Cattle Mastitis

Author: WangYingJie
Tutor: AnLiGuo;ZhongZuoFeng
School: Shandong Normal University
Course: Cell Biology
Keywords: Mastitis in dairy cattle Staphylococcus aureus Coagulase Adhesins Hemolysin Immunity
CLC: S852.61
Type: Master's thesis
Year: 2008
Downloads: 224
Quote: 2
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Abstract


The separation of the experiments carried out on the of Bovine Mastitis positive and negative milk bacteria detected Staphylococcus aureus, Staphylococcus aureus-specific detection primers. Staphylococcus aureus hemolysin is a toxin that is the main one of the virulence factors, including α, β, γ and δ toxin. Hemolysin in a variety of mammalian red blood cell hemolysis, and acting on the smooth muscle cells, causing breast smooth muscle contraction, paralysis, caused by cow's gangrenous mastitis. The experimental design double PCR method to detect Staphylococcus aureus isolated in Shandong province, more than 20 large-scale dairy mastitis cases was found: more than 80% of S. aureus alpha hemolysin gene exists, 704bp gene fragment ; hemolysin gene and β ratio of 59%, fragment size 496bp. Detection of S. aureus the adhesion gene Fnbp A Fnbp B, analysis Bovine Mastitis Staphylococcus aureus clinical isolates fibronectin protein connexin major gene. The use of design and synthesis of specific primers for PCR amplification of Staphylococcus aureus the double PCR detection of Staphylococcus aureus clinical isolates isolated and identified in the laboratory. PCR products after electrophoresis imaging showed a specific band of 524bp and 642bp, respectively. More than 95% of the S. aureus found that by double PCR testing on clinical isolates of Staphylococcus aureus: presence of the the adhesin gene Fnbp A, while only 10% Fnbp B gene. Coagulase important indicator for testing the pathogenicity of Staphylococcus aureus, which enables the anticoagulant-containing rabbit or human plasma coagulation, help pathogenic resist the role of the host body phagocytic cells and bactericidal substances. Basis Staphylococcus aureus plasma coagulase gene sequences, specific primers were designed and synthesized, and its gene was amplified by polymerase chain reaction and amplified gene was cloned into T vector for sequencing, the separation of cows mastitis pathogens were identified. The results show: Staphylococcus aureus amplified specific band size of 761bp; other strain does not appear to strip. The Staphylococcus aureus Fnbp the antibody not only can prevent bacterial adhesion, but also a opsonin, can promote the body's bacterial clearance. Staphylococcus aureus, the α-hemolysin is an extracellular toxin, and its toxoid sheep caused by Staphylococcus aureus mastitis partial protection. Obtained in the experiment using genetically engineered proteins rFnbp of rα-HL and eukaryotic expression plasmid with pcDNA3.1-ClfA mixed adjuvant laboratory mice, detected by ELISA in serum antibody levels. The results showed that: All protein vaccine experiment mice were collected after the first free serum specific antibodies were detected by preliminary Staphylococcus aureus challenge experiments confirmed genetic engineering subunit vaccines made of mice Staphylococcus aureus infection and mortality, provide a new avenue for the prevention and treatment of Staphylococcus aureus.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Basic Veterinary Science > Animal Microbiology ( Veterinary Microbiology, ) > Pathogenic bacteria
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