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AB5075, a Highly Virulent Isolate of Acinetobacter baumannii, as a Model Strain for the Evaluation of Pathogenesis and Antimicrobial Treatments

机译：AB5075，鲍曼不动杆菌的高毒力分离株，是用于评估发病机理和抗菌治疗的模型菌株

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Acinetobacter baumannii is recognized as an emerging bacterial pathogen because of traits such as prolonged survival in a desiccated state, effective nosocomial transmission, and an inherent ability to acquire antibiotic resistance genes. A pressing need in the field of A.?baumannii research is a suitable model strain that is representative of current clinical isolates, is highly virulent in established animal models, and can be genetically manipulated. To identify a suitable strain, a genetically diverse set of recent U.S. military clinical isolates was assessed. Pulsed-field gel electrophoresis and multiplex PCR determined the genetic diversity of 33 A.?baumannii isolates. Subsequently, five representative isolates were tested in murine pulmonary and Galleria mellonella models of infection. Infections with one strain, AB5075, were considerably more severe in both animal models than those with other isolates, as there was a significant decrease in survival rates. AB5075 also caused osteomyelitis in a rat open fracture model, while another isolate did not. Additionally, a Tn5 transposon library was successfully generated in AB5075, and the insertion of exogenous genes into the AB5075 chromosome via Tn7 was completed, suggesting that this isolate may be genetically amenable for research purposes. Finally, proof-of-concept experiments with the antibiotic rifampin showed that this strain can be used in animal models to assess therapies under numerous parameters, including survival rates and lung bacterial burden. We propose that AB5075 can serve as a model strain for A.?baumannii pathogenesis due to its relatively recent isolation, multidrug resistance, reproducible virulence in animal models, and genetic tractability. >IMPORTANCE The incidence of A.?baumannii infections has increased over the last decade, and unfortunately, so has antibiotic resistance in this bacterial species. A.?baumannii is now responsible for more than 10% of all hospital-acquired infections in the United States and has a >50% mortality rate in patients with sepsis and pneumonia. Most research on the pathogenicity of A.?baumannii focused on isolates that are not truly representative of current multidrug-resistant strains isolated from patients. After screening of a panel of isolates in different in vitro and in vivo assays, the strain AB5075 was selected as more suitable for research because of its antibiotic resistance profile and increased virulence in animal models. Moreover, AB5075 is susceptible to tetracycline and hygromycin, which makes it amenable to genetic manipulation. Taken together, these traits make AB5075 a good candidate for use in studying virulence and pathogenicity of this species and testing novel antimicrobials.
机译：鲍曼不动杆菌被认为是一种新兴的细菌病原体，其特征包括在干燥状态下可延长生存时间，有效的医院内传播以及获得抗生素抗性基因的固有能力。鲍曼不动杆菌的研究领域中迫切需要一种合适的模型菌株，该菌株可代表当前的临床分离株，在已建立的动物模型中具有高毒性，并且可以进行基因操作。为了鉴定合适的菌株，评估了遗传多样性的一组最近的美国军事临床分离株。脉冲场凝胶电泳和多重PCR确定了33株鲍曼不动杆菌的遗传多样性。随后，在鼠的肺部感染和梅勒氏菌感染模型中测试了五种代表性分离株。在两种动物模型中，一种菌株AB5075的感染都比其他分离株的感染严重得多，因为存活率显着降低。 AB5075还可以在大鼠开放性骨折模型中引起骨髓炎，而另一种分离物则没有。此外，在AB5075中成功生成了一个Tn 5 转座子文库，并通过Tn 7 完成了将外源基因插入AB5075染色体，这表明该分离株可能是遗传性的适合研究目的。最后，使用抗生素利福平进行的概念验证实验表明，该菌株可用于动物模型，以评估包括存活率和肺部细菌负担在内的许多参数下的治疗方法。我们建议AB5075可以作为鲍氏不动杆菌的发病机制的模型菌株，因为它具有相对较近的分离性，多药耐药性，动物模型中的可复制毒力以及遗传易处理性。 >重要性在过去十年中，鲍曼不动杆菌的感染率有所增加，不幸的是，这种细菌的耐药性也有所增加。在美国，鲍曼不动杆菌现在占所有医院获得性感染的10％以上，败血症和肺炎患者的死亡率超过50％。关于鲍曼不动杆菌的致病性的大多数研究都集中在不能真正代表目前从患者身上分离出来的多药耐药菌株的分离株上。在通过不同的体外和体内分析方法筛选出一组分离株后，AB5075菌株被选为更适合研究的菌株，因为它具有抗药性和高毒力。动物模型。此外，AB5075对四环素和潮霉素敏感，这使其易于进行基因操作。综上所述，这些特性使AB5075成为用于研究该物种的毒力和致病性并测试新型抗菌剂的良好候选者。

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