肺炎克雷伯菌不仅是一种世界性的人类病原菌,它也影响野生动物，例如大熊猫（Ailuropodamelanoleuca），最近被证明会导致腹泻，器官衰竭,甚至死亡。2018年在成都大熊猫繁育研究基地进行了肺炎克雷伯菌调查。作为调查的一部分,脉冲场凝胶电泳（PFGE）分型，多位点序列分型（MLST），抗生素耐药性谱(ARPs)，并在所有分离株的基础上研究了抗生素抗性基因(ARGs)。2018年5月至12月，从72个黑藻中收集粪便样本，从153个粪便样本中分离出90个肺炎克雷伯菌。基因分型结果表明,分离株具有较高的多样性,其中PFGE获得84个簇，MLST获得57个簇。分离株相似度的总体趋势是第一样本期>第二样本期>第三样本期，这表明肺炎克雷伯菌基因组变异性的增加。此外,90株对氨苄青霉素有较高的耐药性,利福平,和复方磺胺甲恶唑。在获得的分离物中，50%携带6~8个ARPs，在三个采样周期内，载运量增加，其中我们发现了两个在第三个样本期间携带12和13个ARP的分离株，分别。此外，共检测到65种ARGs(90.28%，65/72）在90例肺炎克雷伯菌样本中。几乎所有取样的细菌都含有17种属于β-内酰胺酶的ARGs，多药，MGEs,氨基糖苷类,和四环素,这可能是肺炎克雷伯菌ARP的基础。此外，多药和MGE的类型对肺炎克雷伯菌的抗生素敏感性有较大影响.我们的结果表明，肺炎克雷伯菌在黑藻中具有严重的传播风险，肺炎克雷伯菌在大量抗生素使用下具有很高的基因组多样性和药物耐受性风险。
Klebsiella pneumoniae is not only a worldwide human pathogen, it also effects wildlife, such as the giant panda (Ailuropoda melanoleuca), in which it has recently been evidenced to result in diarrhea, organ failure, and even death. A K. pneumoniae investigation was carried out at the Chengdu Research Base of Giant Panda Breeding in 2018. As part of the investigation, the pulsed-field gel electrophoresis (PFGE) typing, multilocus-sequence typing (MLST), antibiotic resistance profiles (ARPs), and antibiotic resistance genes (ARGs) were studied based on all isolates. Fecal samples were collected from 72 A. melanoleuca from May to December 2018, and a total of 90 K. pneumoniae were isolated from 153 fecal samples. The genotyping results showed that the isolates had high diversity, of which 84 clusters were obtained by PFGE and 57 STs by MLST. The overall trend of the similarity of isolates was the first sample period > second sample period > third sample period, which showed the increasement of genome variability of K. pneumoniae. In addition, 90 isolates showed high resistance to ampicillin, rifampicin, and compound sulfamethoxazole. Of the obtained isolates, 50% carried 6~8 ARPs, and the carrying volume increased during three sample periods, in which we found two isolates carrying 12 and 13 ARPs during the third sample period, respectively. Moreover, a total of 65 ARGs were detected (90.28%, 65/72) in 90 K. pneumoniae samples. Almost all bacteria sampled contained 17 ARGs that belonged to the β-lactamase, Multidrug, MGEs, Aminoglycoside, and Tetracycline, which may be the basis of ARPs of K. pneumoniae. Moreover, the types of Multidrug and MGEs had a greater impact on antibiotic susceptivity of K. pneumoniae. Our results showed that K. pneumoniae has a serious risk of transmission in A. melanoleuca and K. pneumoniae had a high possibility of genome diversity and the risk of drugs tolerance under the large antibiotic usage.
