1株猪源ST-35型肺炎克雷伯菌的致病性和药物敏感性分析

doi:10.11843/j.issn.0366-6964.2022.12.021

摘要/Abstract

摘要： 旨在了解从猪肺组织中分离得到的肺炎克雷伯菌的致病性和药物敏感性，本研究通过形态学鉴定、PCR鉴定、16S rRNA基因测序和MLST多位点序列分析等方法对分离菌株进行鉴定，毒力基因检测和小鼠攻毒试验确定分离株的致病性，并采用抗生素和中药分析药物敏感性。结果表明，分离菌株为革兰阴性杆菌，生化结果与肺炎克雷伯菌生化特性相符；16S rRNA基因序列比对分析和系统发育树构建确定分离菌株为肺炎克雷伯菌，命名为肺炎克雷伯菌KP-0728；MLST分析显示，KP-0728属于ST-35型，与ST-875汇聚在一支；KP-0728含有uge、wabG、fimH、kfu 4种毒力基因，感染小鼠可导致小鼠多个器官不同程度的病变，高剂量感染可导致小鼠死亡；KP-0728携带bla-SHV、sul2、tetA、aadA1 4种耐药基因；KP-0728对氨苄西林、亚胺培南、庆大霉素、四环素、复方新诺明耐药；对头孢曲松、头孢氨苄、头孢美唑、阿奇霉素、氧氟沙星、多黏菌素B、替考拉宁敏感；9味中药体外抑菌试验结果显示，茯苓对肺炎克雷伯菌分离株KP-0728的抑菌效果最为显著。动物治疗试验相关数据显示茯苓对KP-0728具有体内抑制作用。综上，本研究分离到1株猪源ST-35型肺炎克雷伯菌，携带多种毒力基因和耐药基因，人工感染小鼠可致小鼠发病，半数致死量(LD₅₀)为4.56×10⁶ CFU，对中药茯苓和多种抗生素敏感，研究结果为肺炎克雷伯菌的防治和疫苗研究提供了参考依据。

关键词: 肺炎克雷伯菌, 分离鉴定, 致病性, 耐药性, 中药

Abstract: This study was conducted to understand the pathogenicity and drug sensitivity of Klebsiella pneumoniae (Kp) isolated from pig lung tissue. In this study, the isolate was identified based on morphology observation, PCR detection, 16S rRNA sequencing and MLST multi-locus sequence analysis. The pathogenicity of the isolate was determined by virulence gene detection and mouse challenge test, and the drug sensitivity was analyzed by antibiotics and traditional Chinese medicine. The results showed that the isolate was Gram-negative bacilli, and the biochemical results were consistent with the biochemical characteristics of Kp. 16S rRNA gene sequence alignment analysis and phylogenetic tree construction identified the isolated strain as Kp, named KP-0728. MLST analysis showed that KP-0728 belonged to ST-35 type and converged with ST-875. KP-0728 contained four virulence genes, uge, wabG, fimH and kfu. When KP-0728 was infected with mice intraperitoneally, different degrees of pathological changes in various organs of the infected mice were induced, and high dose infection of KP-0728 can cause death in mice. KP-0728 carried four resistance genes bla-SHV, sul2, tetA and aadA1, and was resistant to ampicillin, imipenem, gentamicin, tetracycline and cotrimoxazole and sensitive to ceftriaxone, cefalexin, cefmetazole, azithromycin, ofloxacin, polymyxin B and teicoplanin. The results of in vitro antibacterial experiments of nine Chinese medicines showed that Poria cocos had the most significant antibacterial effect on KP-0728. The data of animal treatment test also showed that Poria cocos had inhibitory effect on KP-0728 in vivo. In conclusion, a porcine Kp type ST-35 was isolated in this study, carrying a variety of virulence genes and drug resistance genes. Artificial infection of Kp in mice can lead to pathological changes and the median lethal dose (LD₅₀) was 4.56×10⁶ CFU, and KP-0728 is sensitive to traditional Chinese medicine Poria cocos and many kinds of antibiotics. The results provide a reference for the prevention and treatment of Kp and vaccine research.

Key words: Klebsiella pneumoniae, isolation and identification, pathogenicity, drug resistance, traditional Chinese medicine

中图分类号:

S852.61

李华明, 项维, 卢文兵, 刘峰, 雷连成, 张付贤. 1株猪源ST-35型肺炎克雷伯菌的致病性和药物敏感性分析[J]. 畜牧兽医学报, 2022, 53(12): 4356-4366.

LI Huaming, XIANG Wei, LU Wenbing, LIU Feng, LEI Liancheng, ZHANG Fuxian. Pathogenicity and Drug Sensitivity Analysis of a Porcine Klebsiella pneumoniae Type ST-35[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(12): 4356-4366.

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