朱思逸, 李先平, 宋利琼, 肖玉春, 黄元铭, 储琼芳, 任志鸿. 耐万古霉素肠球菌小鼠感染模型的优化[J]. 疾病监测, 2018, 33(12): 979-984. DOI: 10.3784/j.issn.1003-9961.2018.12.004
引用本文: 朱思逸, 李先平, 宋利琼, 肖玉春, 黄元铭, 储琼芳, 任志鸿. 耐万古霉素肠球菌小鼠感染模型的优化[J]. 疾病监测, 2018, 33(12): 979-984. DOI: 10.3784/j.issn.1003-9961.2018.12.004
Siyi Zhu, Xianping Li, Liqiong Song, Yuchun Xiao, Yuanming Huang, Qiongfang Chu, Zhihong Ren. Optimization of vancomycin resistant Enterococcus infection model in mice[J]. Disease Surveillance, 2018, 33(12): 979-984. DOI: 10.3784/j.issn.1003-9961.2018.12.004
Citation: Siyi Zhu, Xianping Li, Liqiong Song, Yuchun Xiao, Yuanming Huang, Qiongfang Chu, Zhihong Ren. Optimization of vancomycin resistant Enterococcus infection model in mice[J]. Disease Surveillance, 2018, 33(12): 979-984. DOI: 10.3784/j.issn.1003-9961.2018.12.004

耐万古霉素肠球菌小鼠感染模型的优化

Optimization of vancomycin resistant Enterococcus infection model in mice

  • 摘要:
    目的 优化小鼠经口感染耐万古霉素肠球菌(VRE)肠道定植模型,明确不同抗生素预处理方式和感染剂量对VRE在肠道定植的影响。
    方法 C57/6J小鼠随机分为3组,分别为混合抗生素、氨苄西林、万古霉素预处理组。 混合抗生素预处理组:感染前第7天给予小鼠混合抗生素(卡那霉素、庆大霉素、粘菌素、甲硝唑、万古霉素)水溶液;感染前第2天,将混合抗生素水溶液换为万古霉素水溶液,感染前1 d,腹腔注射克林霉素。 氨苄西林与克林霉素预处理组:感染前第7天给予氨苄西林水溶液,感染前1 d,腹腔注射克林霉素。 万古霉素预处理组:感染前第7天给予万古霉素水溶液。第0天,3组小鼠同时进行VRE灌胃处理。 感染后第3天,氨苄西林、万古霉素水溶液改换为正常饮用水,直至实验结束。 在感染后2周内对收取的小鼠粪便活菌计数,观察VRE定植量的变化。
    结果 感染后第1天,VRE在3种抗生素预处理的小鼠肠道中均能成功定植。 在感染后1 ~ 9 d,3种模型的粪便VRE含量在105 ~ 108 cfu/mg。 感染后第9天,3组小鼠粪便中VRE定植量出现下降趋势,第15 天(实验结束)粪便VRE含量降至103 ~ 105 cfu/mg。 另外,感染后第10天氨苄西林处理组中有2只小鼠死亡。 对万古霉素预处理模型给予的2个不同感染剂量,在整个急性感染期两组VRE定植量差异无统计学意义。
    结论 万古霉素预处理的VRE肠道定植模型的稳定性和简洁性优于混合抗生素或氨苄西林预处理的模型。 在万古霉素预处理模型中,VRE可在肠道定植至少15 d,且感染剂量在105 ~ 107 cfu/只间不影响VRE的肠道定植模型的建立。

     

    Abstract:
    Objective To optimize the intestinal colonization model of vancomycin resistant Enterococcus (VRE) in mouse model, and evaluate the effect of antibiotic pretreatment and different infection dosages on the intestinal VRE colonization.
    Methods C57/6J mice were randomly divided into 3 pretreatment groups, which were mixed antibiotic pretreatment group, ampicillin and clindamycin pretreatment group and vancomycin pretreatment group. In mixed pretreatment antibiotic group, the mice were given mix antibiotic solution (kanamycin, gentamicin, colistin, metronidazole, vancomycin) on day 7 before infection; on day 2 before infection, vancomycin solution was given instead of the mixed antibiotic solution, and intraperitoneal injection of clindamycin was given on day 1 before infection. In ampicillin and clindamycin pretreatment group, the mice were given ampicillin solution on day 7 before infection, and intraperitoneal injection of clindamycin was given 1 day before infection. In vancomycin pretreatment group, vancomycin solution was given on day 7 before infection. On day 0, three groups of mice were infected with VRE. On day 3 after infection, vancomycin solution and ampicillin solution were changed to normal drinking water. The collected fecal live bacteria were counted within 2 weeks after infection, and changes of VRE colonization were observed.
    Results Three antibiotic-pretreated mouse models infected with VRE were successfully established, and VRE were successfully colonized on day 1 after infection. Within 1–9 days after infection, the VRE contents in feces of the three models was 105–108cfu/mg. Nine days after infection, the colonization of VRE in the three mouse groups showed downward trends, and the content of VRE in the mouse feces decreased to 103–105cfu/mg on the 15th day. And, we observed that two mice died in the ampicillin group on day 10 after infection. There was no significant difference in VRE colonization between high dosage group and low dosage group during the acute infection period in vancomycin pretreated mice.
    Conclusion The stability and simplicity of vancomycin-pretreated mouse model was superior to those of the other two antibiotic-pretreated mouse models. By the end of the experiment, VRE could successfully colonize in the intestinal tract of the mice for more than 15 days in vancomycin pretreatment group, and the VRE intestinal colonization model could be established steadily at the infection dosages ranging from 105 cfu to 107 cfu per mouse.

     

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