菪可轮蒲水煎液对多杀性巴氏杆菌体内外抑菌活性研究
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摘要
研究菪可轮蒲水煎液对多杀性巴氏杆菌的体内外抑菌活性作用。用传统水煎法制备菪可轮蒲水煎液,用HPLC法测定菪可轮蒲水煎液中龙胆苦苷浓度,并用龙胆苦苷浓度标定复方药物浓度,分别采用牛津杯法和宏量肉汤稀释法测定菪可轮蒲水煎液对多杀性巴氏杆菌的抑菌圈直径和最小抑菌浓度(MIC),通过建立死亡率为75%的多杀性巴氏杆菌体内感染小鼠模型,测定浓度为0.023 2 mg/mL的菪可轮蒲水煎液,在感染前和感染后给药方式下,分别给予0.1、0.3、0.5 mL药物时对小鼠多杀性巴氏杆菌感染的预防和治疗作用。结果显示,菪可轮蒲水煎液浓度为0.046 4 mg/mL时,对多杀性巴氏杆菌的抑菌圈平均直径为13.5 mm,MIC值为0.002 9 mg/mL,经病理剖检、细菌镜检、全自动微生物鉴定及药敏系统(VITEK 2 COMPACT)细菌鉴定,表明死亡率为75%的多杀性巴氏杆菌体内感染小鼠模型建立成功,且预防组各组和治疗组各组小鼠死亡率依次为58.3%、41.7%、83.3%、66.7%、58.3%和91.7%。综合以上试验结果,菪可轮蒲水煎液对多杀性巴氏杆菌具有良好的体内外抑菌活性,且其预防作用较治疗作用明显。
To study the antibacterial effect of Dang-Ke-Lun-Pu water extract on Pasteurella multocda in vitro and in vivo,the effective components were extracted with water-extracting in the compound,the content of Gentiopicroside in Dang-Ke-Lun-Pu water extract was measured by HPLC method,and its content indicates the drug concentration of Dang-Ke-Lun-Pu water extract.The antibacterial ring diameters and minimal inhibitory concentration(MIC) were determined by Oxford plate assay system and a broth macro dilution assay.The model of Pasteurella multocida infection with a mortality rate of 75% in mice were established.The prevention and treatment effects of mice of different doses with 0.1 mL,0.3 mL,0.5 mL respectively were measured when the compound concentration is 0.023 2 mg/mL before and after infection.When Dang-Ke-Lun-Pu water extract concentration is 0.046 4 mg/mL,the average diameter of antibacterial rings of water extract against Pasteurella multocida was 13.5 mm, and the MIC was 0.002 9 mg/mL.Pathological examination,bacterial microscopy,VITEK 2 COMPACT identification after infection showed that the mice infected with Pasteurella multocida model was successfully established.The mortality rates of the mice in the prevention group and the treatment group were 58.3%,41.7%,83.3%,66.7%,58.3% and 91.7%,respectively.Based on the above results,Dang-Ke-Lun-Pu water extract has a good antibacterial activity in vitro and in vivo in prevention and treatment Pasteurella multocida infection,and the preventive effect was more significant than the treatment effect.
To study the antibacterial effect of Dang-Ke-Lun-Pu water extract on Pasteurella multocda in vitro and in vivo,the effective components were extracted with water-extracting in the compound,the content of Gentiopicroside in Dang-Ke-Lun-Pu water extract was measured by HPLC method,and its content indicates the drug concentration of Dang-Ke-Lun-Pu water extract.The antibacterial ring diameters and minimal inhibitory concentration(MIC) were determined by Oxford plate assay system and a broth macro dilution assay.The model of Pasteurella multocida infection with a mortality rate of 75% in mice were established.The prevention and treatment effects of mice of different doses with 0.1 mL,0.3 mL,0.5 mL respectively were measured when the compound concentration is 0.023 2 mg/mL before and after infection.When Dang-Ke-Lun-Pu water extract concentration is 0.046 4 mg/mL,the average diameter of antibacterial rings of water extract against Pasteurella multocida was 13.5 mm, and the MIC was 0.002 9 mg/mL.Pathological examination,bacterial microscopy,VITEK 2 COMPACT identification after infection showed that the mice infected with Pasteurella multocida model was successfully established.The mortality rates of the mice in the prevention group and the treatment group were 58.3%,41.7%,83.3%,66.7%,58.3% and 91.7%,respectively.Based on the above results,Dang-Ke-Lun-Pu water extract has a good antibacterial activity in vitro and in vivo in prevention and treatment Pasteurella multocida infection,and the preventive effect was more significant than the treatment effect.
引文
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[14] Du H,Wu C,Li C,et al.Two novel cross protective antigens for bovine Pasteurella multocida[J].Mol Med Rep,2017,16(4):4627-4633.
[15] 牛思博,姜志刚,德艳艳,等.重组牛源荚膜A型多杀性巴氏杆菌脂蛋白E的免疫保护性研究[J].中国预防兽医学报,2013,35(5):401-404.
[16] 白莎,倪健,刘英英,等.高效液相色谱法测定抗风湿颗粒中龙胆苦苷的含量[J].北京中医药大学学报,2004,27(6):61-62.
[17] 卢燕.马波沙星在多杀性巴氏杆菌感染小鼠体内的药动——药效学同步模型研究[D].广东广州:华南农业大学,2016.
[18] 曾庆林,苏佳,万鹏,等.马波沙星在猪多杀性巴氏杆菌感染组织液中药代动力学的研究[J].中国兽医科学,2017,47(10):1326-1330.
[2] Zamrisaad M,Annas S.Vaccination against hemorrhagic septicemia of bovines:a review.[J].Pakistan Vet J,2016,36(1):1-5.
[3] 杨立英.猪肺疫的鉴别诊断及防治措施[J].中国畜牧兽医文摘,2016,32(2):196.
[4] 白挨泉,刘为民,徐苏标,等.广东地区猪传染性萎缩性鼻炎流行病学调查[J].中国预防兽医学报,2012,34(6):456-459.
[5] Islam M T,Ali M H,Chandra A,et al.Standardization of effective dose of fowl cholera vaccine in pigeon in bangladesh [J].Bangladesh J Vet Med,2017,15 (2):97-105.
[6] Chengappa M M,Myers R C,Carter G R.A streptomycin dependent live Pasteurella multocida vaccine for the prevention of rabbit pasteurellosis[J].Lab An Sci,1980,30(3):515.
[7] 马斯真,金玉珍,马仲礼.羊巴氏杆菌病[J].畜牧兽医杂志,2015,34(1):159.
[8] 彭忠,王庚,吴广敬,等.猪源多杀性巴氏杆菌的分离与鉴定[J].中国兽医科学,2015,45(9):943-951.
[9] 潘洪珠,贺洪强,姜涤非.毛皮动物巴氏杆菌病经破损皮肤感染人的病例报告[J].毛皮动物饲养,1990(2):38-39.
[10] 何贤发,冯方东,徐一凡,等.禽源多杀性巴氏杆菌毒力基因多重PCR检测方法的建立[J].动物医学进展,2018,39(4):13-18.
[11] 孔令聪,战利,赵晴,等.动物源多杀性巴氏杆菌耐药性及耐药机制研究进展[J].中国兽药杂志,2012,46(3):52-55.
[12] 孙泰俊,樊海岩.论藏药材资源保护发展的现状和建议[J].中国民族医药杂志,2013,19(7):7-11.
[13] 彭忠,梁婉,吴斌.多杀性巴氏杆菌分子分型方法简述[J].微生物学报,2016,56(10):1521-1529.
[14] Du H,Wu C,Li C,et al.Two novel cross protective antigens for bovine Pasteurella multocida[J].Mol Med Rep,2017,16(4):4627-4633.
[15] 牛思博,姜志刚,德艳艳,等.重组牛源荚膜A型多杀性巴氏杆菌脂蛋白E的免疫保护性研究[J].中国预防兽医学报,2013,35(5):401-404.
[16] 白莎,倪健,刘英英,等.高效液相色谱法测定抗风湿颗粒中龙胆苦苷的含量[J].北京中医药大学学报,2004,27(6):61-62.
[17] 卢燕.马波沙星在多杀性巴氏杆菌感染小鼠体内的药动——药效学同步模型研究[D].广东广州:华南农业大学,2016.
[18] 曾庆林,苏佳,万鹏,等.马波沙星在猪多杀性巴氏杆菌感染组织液中药代动力学的研究[J].中国兽医科学,2017,47(10):1326-1330.