本草乳炎清散对奶牛乳房炎的药效学研究

发布时间：2018-11-08 15:38

【摘要】：本草乳炎清散是内蒙古华天制药有限公司自主研发的兽用天然药物复方制剂,具有清热解毒,消痈散结,消肿止痛,托毒排脓的功效。目的:通过进行奶牛乳房炎病原菌分离鉴定、本草乳炎清散体外抑菌、抗炎、急性毒性和临床治疗试验,评价本草乳炎清散针对奶牛乳房炎的临床治疗效果及其对机体毒性强弱,为本草乳炎清散的临床应用提供科学依据。方法:(1)试验运用小鼠耳廓肿胀模型、皮下棉球炎性肉芽肿增生模型和腹腔内血管在醋酸刺激下通透性升高模型,观察本草乳炎清散对炎症的消除作用;(2)通过对乳样进行细菌分离纯化,利用染色、镜检进行形态学观察以及生化实验鉴定乳样中的病原菌;(3)选择牛津小杯法检测本草乳炎清散对奶牛乳房炎主要致病菌的抑菌圈大小;试管倍比稀释法确定最小抑菌浓度(MIC)和最小杀菌浓度(MBC);(4)选择测定小鼠对本草乳炎清散的最大耐受量(MTD)和最大耐受量倍数进行安全性评价;(5)经混饲投喂本草乳炎清散,通过临床治愈率、有效率,评价其对奶牛乳房炎的临床治疗效果。结果:(1)抗炎试验:本草乳炎清散高、中、低剂量组耳廓肿胀抑制率分别为39.74%、36.78%和28.84%,与生理盐水对照组相比,高、中剂量组存在极显著性差异(P0.01),低剂量组存在显著性差异(P0.05);本草乳炎清散高、中、低剂量组小鼠肉芽肿增生抑制率分别为53.53%、51.69%和35.89%,与生理盐水对照组相比,高、中、低剂量组均存在极显著性差异(P0.01);本草乳炎清散高、中、低剂量组对小鼠腹腔毛细血管通透性升高抑制率分别为51.06%、46.81%和34.04%,与生理盐水对照组相比,高、中剂量组存在极显著性差异(P0.01),低剂量组存在显著性差异(P0.05);(2)细菌分离鉴定:共得到141株细菌,4个种类:无乳链球菌、金黄色葡萄球菌、大肠埃希菌和表皮葡萄球菌,检出率分别为:46.10%、24.11%、20.57%和9.22%;(3)体外抑菌试验:本草乳炎清散对金黄色葡萄球菌、大肠埃希菌、无乳链球菌的MIC分别为0.0625、0.125、0.0625g·mL-1;MBC分别为0.125、0.250、0.125g·mL-1。本草乳炎清散(0.50g·mL-1)对金黄色葡萄球菌、大肠埃希菌、无乳链球菌的抑菌圈直径分别为22.83、21.60、24.51mm,均大于20mm,极敏感。(4)急性毒性试验:本草乳炎清散对小鼠的最大耐受量为600mg/10g BW,最大耐受量倍数为150倍,属于实际无毒,安全性较高;(5)临床治疗试验:本草乳炎清散高、中剂量组治疗有效率均达100%,且低剂量组的有效率高于公英散对照组。结论:本草乳炎清散对炎症有很好的消除作用,对奶牛乳房炎主要致病菌的抑制效果明显,在正常使用剂量内安全性好,能够有效治疗对奶牛乳房炎。
[Abstract]:Ru Yan Qing San is a natural veterinary medicine compound preparation independently developed by Inner Mongolia Huatian Pharmaceutical Co., Ltd. It has the effect of clearing heat and detoxification, eliminating carbuncle, eliminating swelling and relieving pain, holding poison and discharging pus. Objective: to isolate and identify the pathogenic bacteria of dairy cow mastitis, and to investigate the effect of Ruyangqing powder on the inhibition, anti-inflammatory, acute toxicity and clinical treatment of dairy cow mastitis in vitro. To evaluate the clinical therapeutic effect of Bencao Ruyanqing Powder (BRYQS) on dairy cow mastitis and to provide scientific basis for clinical application of Bencao Ruyan Qingsan. Methods: (1) the mice auricle swelling model, subcutaneous cotton ball inflammatory granulomatous hyperplasia model and the model of increased permeability of intraperitoneal blood vessels stimulated by acetic acid were used to observe the anti-inflammatory effect of Bencao Ruyanqing Powder. (2) the bacteria in milk samples were isolated and purified, the pathogens in milk samples were identified by staining, microscopic observation and biochemical experiments. (3) the inhibition circle size of the main pathogenic bacteria of dairy cow mastitis was detected by Oxford small cup method, and the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC);) were determined by tube doubling dilution method. (4) to evaluate the safety of the maximum tolerance dose (MTD) and the maximum tolerance dose (multiple) of Ruyanqing Powder in mice. (5) to evaluate the clinical treatment of dairy cow mastitis through the clinical cure rate and effective rate. Results: (1) Anti-inflammatory test: the inhibition rates of auricle swelling in the high, middle and low dose groups were 39.74% and 28.84%, respectively, which were higher than those in the normal saline control group. There was significant difference in middle dose group (P0.01) and low dose group (P0.05). The inhibition rates of granulomatous proliferation were 53.53% and 35.89% in the high, medium and low dose groups, respectively. Compared with the normal saline control group, the inhibition rates of granulomatous proliferation were significantly higher (P0.01). The inhibition rates of high, medium and low dose groups on the increase of celiac capillary permeability in mice were 51.06% and 34.04%, respectively, which were higher than those in normal saline group. There was significant difference in middle dose group (P0.01) and low dose group (P0.05). (2) bacterial isolation and identification: a total of 141 strains of bacteria were obtained, four species: Streptococcus lactococcus, Staphylococcus aureus, Escherichia coli and Staphylococcus epidermidis, the detection rates were: 46.1010, 24.11, 20.57% and 9.22, respectively; (3) in vitro bacteriostasis test: the MIC of Ruyangqing Powder against Staphylococcus aureus, Escherichia coli and Streptococcus lactococcus were 0.06250.125 ~ 0.0625g mL-1;MBC = 0.1250.250,0.125g mL-1., respectively. The bacteriostasis diameter of RYQS (0.50g mL-1) against Staphylococcus aureus, Escherichia coli and Streptococcus lactobacillus were 22.83 ~ 21.60 ~ 24.51 mm, > 20mm, respectively. (4) Acute toxicity test: the maximum tolerance dose of RUYQS to mice was 150 times that of 600mg/10g BW, which was non-toxic and safe. (5) Clinical treatment trial: the effective rate of low dose group was higher than that of Gongying powder group, the effective rate of middle dose group was 100 kum, and the effective rate of low dose group was higher than that of Gongying powder group. Conclusion: Bencao Ruyanqing Powder has a good effect on the elimination of inflammation and has obvious inhibitory effect on the main pathogenic bacteria of mastitis in dairy cattle. It is safe in the normal dosage and can effectively treat the mastitis of dairy cow.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S858.23

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