小鼠嗜肺巴斯德杆菌检测方法优化及感染小鼠的药物净化

doi:10.12300/j.issn.1674-5817.2020.197

实验动物与比较医学 ›› 2021, Vol. 41 ›› Issue (4): 351-357.DOI: 10.12300/j.issn.1674-5817.2020.197

小鼠嗜肺巴斯德杆菌检测方法优化及感染小鼠的药物净化

张韬^1,*, 崔璨^2,*, 马贯中¹, 张爱华¹

1.南京医科大学医药实验动物中心,南京 211166;
2.上海南方模式生物科技股份有限公司,上海 200120

收稿日期:2020-11-19 修回日期:2021-03-31 发布日期:2021-08-30
作者简介:张韬(1989—), 女, 硕士, 助理实验师, 研究方向: 实验动物质量控制。E-mail: zhangtao@njmu.edu.cn; 崔璨(1989— ), 男, 硕士, 研究方向: 预防兽医。E-mail: 779349408@qq.com; ^*共同第一作者
基金资助:
南京医科大学科技发展基金资助项目(2017NJMU023)

Improvement of Detection Method of Pasteurella pneumotropica and Its Medical Elimination in Mice

ZHANG Tao^1,*, CUI Can^2,*, MA Guanzhong¹, ZHANG Aihua¹

1. Animal Core Facility of Nanjing Medical University, Nanjing 211166, China;
2. Shanghai Model Organisms Center, Inc., Shanghai 200120, China

Received:2020-11-19 Revised:2021-03-31 Published:2021-08-30

摘要/Abstract

摘要： 目的优化小鼠嗜肺巴斯德杆菌（Pasteurella pneumotropica）检测方法,提高检测敏感性以强化嗜肺巴斯德杆菌的日常监测,并探讨感染小鼠药物净化的可行性。方法采用直接涂板法和改良增菌法同时检测嗜肺巴斯德杆菌,比较两种方法的检测效果。采用纸片扩散法研究本地分离的嗜肺巴斯德杆菌菌株对12种抗生素的敏感性。48只雌性C57BL/6小鼠随机分为6组,每组8只,用不同质量浓度的恩诺沙星连续给药6周,评估恩诺沙星在C57BL/6小鼠的安全使用剂量。用本地分离的嗜肺巴斯德杆菌感染小鼠,然后给予嗜肺巴斯德杆菌阳性鼠以含不同质量浓度恩诺沙星的饮用水,检测恩诺沙星对嗜肺巴斯德杆菌感染小鼠的清除效果,确定药物净化方案。结果直接涂板法和改良增菌法检测嗜肺巴斯德杆菌的检出率分别为1.3%和15.1%（P<0.05）;嗜肺巴斯德杆菌对恩诺沙星、头孢西丁和头孢羟唑等抗生素敏感;300 mg·kg^–1·d^–1以下剂量的恩诺沙星饮水给药对小鼠无明显毒副作用;85 mg·kg^–1·d^–1以上剂量的恩诺沙星对嗜肺巴斯德杆菌阳性成年鼠饮用水给药,用药2周后均转阴,停药6周仍未转阳。结论改良增菌法检测嗜肺巴斯德杆菌的敏感性高于直接涂板法,且流程简单便捷,适用于动物房的日常健康监测。85 mg·kg^–1·d^–1以上剂量的恩诺沙星饮水给药能安全有效地清除小鼠嗜肺巴斯德杆菌,必要时可以作为嗜肺巴斯德杆菌感染小鼠的应急净化方案。

关键词: 嗜肺巴斯德杆菌, 检测方法优化, 恩诺沙星, 药敏实验, 药物净化

Abstract: Objective To modify the method for sensitive detection of Pasteurella pneumotropica in mice in routine monitoring, and to explore the feasibility of medical elimination in Pasteurella pneumotropica infected mice.Methods The Pasteurella pneumotropica were detected by routine procedure and modified method that using fluid medium before inoculation. The detection rates of the two methods were compared in statistics. The sensitivity of Pasteurella pneumotropica isolated from local facility to 12 antibiotics was studied by antimicrobial disk sensitivity test. Forty-eight female C57BL/6 mice were randomly divided into 6 groups, with 8 in each group. All the groups were set to different concentration of enrofloxacin continuously for 6 weeks to find out the safe dosage. Mice were infected with local Pasteurella pneumotropica strain, the Pasteurella pneumotropica positive mice were administered enrofloxacin in drinking water at different doses to evaluate the effect of Pasteurella pneumotropica clearance results.Results The detection rates of Pasteurella pneumotropica by routine procedure and improved method were 1.3% and 15.1%, respectively. Pasteurella pneumotropica was sensitive to enrofloxacin, cefoxitin and cefadrazole etc. The mice were safe to enrofloxacin within a daily dosage of 300 mg/kg. The Pasteurella pneumotropica was not detected from positive mice treated with 85 mg/kg enrofloxacin after 2 weeks, and kept negative after withdrawl for 6 weeks.Conclusion Using fluid medium before inoculation can improve the positive detection rate of Pasteurella pneumotropica, which is simple, convenient, and suitable for daily health monitoring of animal facility. Oral administration of enrofloxacin more than a dosage of 85 mg·kg^–1·d^–1 can elimilate Pasteurella pneumotropica from the positive mice safely and effectively, and be an alternative contingency plan for the elimination of Pasteurella pneumotropica in mice.

Key words: Pasteurella pneumotropica, Detection method improvement, Enrofloxacin, Antimicrobial susceptibility test, Medical elimination

中图分类号:

Q95-33
R-332

张韬, 崔璨, 马贯中, 张爱华. 小鼠嗜肺巴斯德杆菌检测方法优化及感染小鼠的药物净化[J]. 实验动物与比较医学, 2021, 41(4): 351-357.

ZHANG Tao, CUI Can, MA Guanzhong, ZHANG Aihua. Improvement of Detection Method of Pasteurella pneumotropica and Its Medical Elimination in Mice[J]. Laboratory Animal and Comparative Medicine, 2021, 41(4): 351-357.

图/表 5

参考文献

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小鼠嗜肺巴斯德杆菌检测方法优化及感染小鼠的药物净化

Improvement of Detection Method of Pasteurella pneumotropica and Its Medical Elimination in Mice

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