| The incidence of dairy cow mastitis is high,and it causes serious economic harm to dairy farming industry.It was often induced by nipple injury,milk siltation and pathogenic bacteria infection,etc.,the infection rate caused by bacteria and fungi accounted for more than 90%.The first choice of treatment is antibiotics,which has a rapid and significant effect,but the long-term use of antibiotics will lead to the tolerance,and cause antibiotics residues in milk,which may pose a safety hazard to human and livestock health.To this end,based on the green,low toxicity Mongolian medicine developed by the research group which is effective in curing dairy cattle mastitis,we selected the antibacterial and anti-inflammatory immune effective components to replace antibiotics,and induced pathological model of mastitis mice with S.aureus and E.coli mixed bacteria liquid,studied the therapeutic effect of the compound on mastitis and the effect on mammary inflammation model mice immune function.The test method and results are as follows:(1)The concentration of mice mastitis pathological model Escherichia coli and s.aureus mixed bacteria which was screened by gradient bacteria and histopathology was:1 × 108cfu/ml.The model has a complete pathological change process.It's successful in creating the model.(2)Prepare the pathological section by HE staining method.Observe the damage degree of pathogenic bacteria on mammary gland of mice.The thickness of the mammary gland and the width of the stroma were measured by a microscope with micrometer.The number of colonies in the mammary gland homogenate was measured by plate count method.Mongolian medicine compound obviously decreased the thickness of mammary gland acinus and colony colonization quantity in mastitis model mice(P<0.05),significantly improved the interstitial width of mammary gland(P<0.05).It shows that the Mongolian medicine compound has good antibacterial and anti-inflammatory effect,and reduces the damage of mammary gland.(3)Measure mastitis model mice body weight,thymus and spleen wet weight,calculate organ index?The results suggest that Mongolian medicine compound can improve immune organ index to reduce immune injury caused by mastitis.(4)Detect the activity of nuclear macrophage in mastitis model in carbon-wide serum,the results suggest that Mongolian medicine compound signifi antly mprove the phagocytosis function of monocyte macrophage in mastitis model mice.(5)Observe the activity of NK cells in mastitis model mice by using MTT assay,the results suggest that Mongolian medicine compound can signifcatly improve the phagocytosis activity of NK cells in mastitis model mice(6)Detect the percentage of peripheral blood lymphocytes in mastitis model mice by using flow cytometry,the results showed that Mongolian medicine compound significantly increased the level of CD3+,CD4+,CD19 ?in mastitis model mice,and decreased the level of CD8 +(P<0.05).Collect the blood samples and detect the content of immunoglobulin in serum by ELISA.The results suggest that the compound of Mongolian medicine compound significantly improve the serum IgG and IgA in mastitis model mice.Prepare mammary gland tissue homogenate,detect the content of cytokines in breast tissue by ELISA.The results suggest that the content of Mongolian medicine compound significantly increased the level of IL-2?IL-10 and IFN-y(P<0.05),decrease IL-11??IL-4?IL-6?IL-8?TNF-?.Conclusion:The Mongolian medicine compound recipe can enhance non-specific immunity and specific immunity of mice by improving immune organ index,phagocytosis of monocyte macrophages,killing activity of NK cells and stability of Thl and Th2 cells,so as to cure or prevent the mastitis caused by mixed infection of E.coli and S.aureus. |
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