畜牧兽医学报 ›› 2010, Vol. 41 ›› Issue (4): 489-494.doi:

• 基础兽医 • 上一篇    下一篇

黄连汤对鸭疫里默氏菌的抑菌作用及对菌体形态的影响

张先福1,韦强2,鲍国连2*,季权安2,庞安娜2   

  1. 1. 浙江林学院,临安 311300;2. 浙江省农业科学院,杭州 310021
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-04-25 发布日期:2010-04-25

Inhibition Activity of Coptidis rhizoma Dicoction against Riemerella anatipestifer and Its Effects on Morphology of Bacterial Body

ZHANG Xian-fu1, WEI Qiang2, BAO Guo-lian2*, JI Quan-an2, PANG An-na2   

  1. 1. Zhejiang Forestry College, Lin′an 311300, China;2. Zhejiang Academy of Agricultural Sciences, Hangzhou 310021,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-04-25 Published:2010-04-25

摘要: 研究中药复方黄连汤水提醇沉浓缩物(Ethanol extracts of Coptidis rhizoma decoction,EeCrd)对鸭疫里默氏菌(Riemerella anatipestifer,RA)的体外抑菌规律及其对菌体形态的影响。黄连汤经水提醇沉后,减压浓缩,制成含生药176 g·L-1的EeCrd原液。用PBS将原液倍比稀释成88、88×2-1、88×2-2 ……88×2-9g·L-1等11个梯度浓度的药液。用纸片扩散法和液体倍比稀释法研究EeCrd完全抑菌时的最小抑菌浓度和不完全抑菌时的亚抑菌浓度;取88和88×2-3 g·L-1 EeCrd浓度组药液分别作用17和 3、6、17 h后的细菌,用透射电镜观察菌体形态的变化。纸片扩散法和液体倍比稀释法测定的EeCrd完全抑制RA时的最小浓度均为88×2-2 g·L-1,不完全抑菌时亚抑菌浓度为88×2-3 g·L-1;88×2-4 g·L-1以上稀释度EeCrd基本无抑菌活性;50 μL 的88 g·L-1浓度组的EeCrd作用RA 17 h后,RA菌体浓缩,变小,最终死亡;50 μL的88×2-3 g·L-1浓度组的EeCrd作用RA 3、6、17 h后,RA菌体长度变长,细胞壁变薄,菌内颜色变浅,皱褶减少,纹理不均匀,菌体干瘪、折叠、弯曲,最终死亡。EeCrd可能通过破坏RA外膜结构来抑制细菌增殖。

Abstract: To investigate inhibition activity of Coptidis rhizoma dicoction against Riemerella anatipestifer and its effects on morphology of bacterial body, Coptidis rhizoma dicoction were prepared with water extraction and alcohol sedimentation method. After vacuum distillation, Ethanol extracts of Coptidis rhizoma decoction (EeCrd) which contained crude drug 176 g·L-1 was acquired. This extracts was bi-diluted to make graded concentrations which contained crude drugs 88,88×2-1,88×2-2 ……88×2-9 g·L-1, Minimum inhibitory concentration (MIC) and sub-inhibited concentration of EeCrd on Riemeralla anatipestifer in vitro were determined by both disc diffusion test and tube broth assays. Morphology of RA were observed by transmission electron microscope in concentration groups of 88 and 88×2-3 g·L-1 of EeCrd in 17 and 3, 6, 17 h, respectively. MIC of EeCrd on RA is 88×2-2 g·L-1 concentration group by both two methods, sub-inhibited concentration are 88×2-3 g·L-1. Above 88×2-4 g·L-1 concentrations, EeCrd almost have no any antibacterial activities. Under 88 g·L-1 concentrations of EeCrd with 50 μL in 17 h, lengths of RA body concentrated and got shorter, ultimately died. In 88×2-3 g·L-1 of EeCrd in 3, 6 , 17 h, lengths of RA body became longer, cell wall got thinner, the fold inside the body reduced, the texture becomes uneven, finally withered, folded, bent and died. EeCrd achieve the purpose of inhibiting the proliferation of RA by destroying the structure of RA cell wall.