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噬菌体鸡尾酒限制. 的生长。

Phage Cocktails Constrain the Growth of .

机构信息

Department of Molecular Biology and Biochemistry, University of California, Irvinegrid.266093.8, California, USA.

Department of Pathology, University of California, San Diegogrid.266100.3, California, USA.

出版信息

mSystems. 2022 Aug 30;7(4):e0001922. doi: 10.1128/msystems.00019-22. Epub 2022 Jun 28.

Abstract

Phages that infect pathogenic bacteria present a valuable resource for treating antibiotic-resistant infections. We isolated and developed a collection of 19 phages, including myoviruses, siphoviruses, and a podovirus, that can infect both Enterococcus faecalis and Enterococcus faecium. Several of the phages that we found in southern California wastewater were from the subfamily (formerly ) and had a broad host range across both E. faecium and E. faecalis. By searching the NCBI Sequence Read Archive, we showed that these phages are prevalent globally in human and animal microbiomes. is a regular member of healthy human gut microbial communities; however, it is also an opportunistic pathogen responsible for an increasing number of antibiotic-resistant infections. We tested the ability of each phage to clear host cultures and delay the emergence of phage-resistant . We found that some phages were ineffective at clearing cultures individually but were effective when combined into cocktails. Quantitative PCR was used to track phage abundance in cocultures and revealed dynamics ranging from one dominant phage to an even distribution of phage growth. Genomic characterization showed that mutations in exopolysaccharide synthesis genes were consistently found in the presence of phage infection. This work will help to inform cocktail design for , which is an important target for phage therapy applications. Due to the rise in antibiotic resistance, infections are a major health crisis that requires the development of alternative therapies. Phage therapy offers an alternative to antibiotics and has shown promise in both and early clinical studies. Here, we established a collection of 19 phages and tested whether combining phages into cocktails could delay growth and the emergence of resistant mutants in comparison with individual phages. We showed that cocktails of two or three phages often prevented the growth of phage-resistant mutants, and we identified which phages were replicating the most in each cocktail. When resistant mutants emerged to single phages, they showed consistent accumulation of mutations in exopolysaccharide synthesis genes. These data serve to demonstrate that a cocktail approach can inform efforts to improve efficacy against isolates and reduce the emergence of resistance.

摘要

噬菌体能感染致病菌,为治疗抗生素耐药感染提供了宝贵资源。我们分离并开发了一个包含 19 种噬菌体的集合,包括肌尾噬菌体、长尾噬菌体和痘病毒,它们可以感染粪肠球菌和屎肠球菌。我们在加利福尼亚南部废水中发现的一些噬菌体来自肌尾噬菌体亚科(以前称为长尾噬菌体科),对粪肠球菌和屎肠球菌都有广泛的宿主范围。通过在 NCBI 序列读取档案中搜索,我们表明这些噬菌体在全球人类和动物微生物组中普遍存在。是健康人类肠道微生物群落的常规成员;然而,它也是一种机会性病原体,导致越来越多的抗生素耐药感染。我们测试了每种噬菌体清除宿主培养物并延迟噬菌体耐药出现的能力。我们发现,一些噬菌体单独清除培养物的效果不佳,但组合成鸡尾酒时效果更好。定量 PCR 用于跟踪共培养物中的噬菌体丰度,并揭示了从一种优势噬菌体到噬菌体生长均匀分布的动态变化。基因组特征表明,在噬菌体感染存在的情况下,始终会发现多糖合成基因的突变。这项工作将有助于为提供鸡尾酒设计信息,这是噬菌体治疗应用的重要目标。由于抗生素耐药性的增加,感染是一个重大的健康危机,需要开发替代疗法。噬菌体治疗为抗生素提供了替代方案,并在和早期临床研究中显示出前景。在这里,我们建立了一个包含 19 种噬菌体的集合,并测试了将噬菌体组合成鸡尾酒是否可以与单独使用噬菌体相比延迟生长和耐药突变体的出现。我们表明,两种或三种噬菌体的鸡尾酒通常可以防止噬菌体耐药突变体的生长,并且我们确定了每种鸡尾酒中哪种噬菌体复制最多。当对单一噬菌体出现耐药突变体时,它们在多糖合成基因中表现出一致的积累突变。这些数据表明鸡尾酒方法可以为提高对抗分离株的疗效和减少耐药性的出现提供信息。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d661/9426582/7713138aff5e/msystems.00019-22-f001.jpg

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