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一株具有显著体内治疗潜力和更高抗铜绿假单胞菌生物膜活性的新型裂解噬菌体。

A novel lytic phage exhibiting a remarkable in vivo therapeutic potential and higher antibiofilm activity against Pseudomonas aeruginosa.

机构信息

Department of Microbiology and Immunology, Faculty of Pharmacy, Zagazig University, Zagazig, 44519, Egypt.

出版信息

Eur J Clin Microbiol Infect Dis. 2023 Oct;42(10):1207-1234. doi: 10.1007/s10096-023-04649-y. Epub 2023 Aug 23.

Abstract

BACKGROUND

Pseudomonas aeruginosa is a nosocomial bacterium responsible for variety of infections. Inappropriate use of antibiotics could lead to emergence of multidrug-resistant (MDR) P. aeruginosa strains. Herein, a virulent phage; vB_PaeM_PS3 was isolated and tested for its application as alternative to antibiotics for controlling P. aeruginosa infections.

METHODS

Phage morphology was observed using transmission electron microscopy (TEM). The phage host range and efficiency of plating (EOP) in addition to phage stability were analyzed. One-step growth curve was performed to detect phage growth kinetics. The impact of isolated phage on planktonic cells and biofilms was assessed. The phage genome was sequenced. Finally, the therapeutic potential of vB_PaeM_PS3 was determined in vivo.

RESULTS

Isolated phage has an icosahedral head and a contractile tail and was assigned to the family Myoviridae. The phage vB_PaeM_PS3 displayed a broad host range, strong bacteriolytic ability, and higher environmental stability. Isolated phage showed a short latent period and large burst size. Importantly, the phage vB_PaeM_PS3 effectively eradicated bacterial biofilms. The genome of vB_PaeM_PS3 consists of 93,922 bp of dsDNA with 49.39% G + C content. It contains 171 predicted open reading frames (ORFs) and 14 genes as tRNA. Interestingly, the phage vB_PaeM_PS3 significantly attenuated P. aeruginosa virulence in host where the survival of bacteria-infected mice was markedly enhanced following phage treatment. Moreover, the colonizing capability of P. aeruginosa was markedly impaired in phage-treated mice as compared to untreated infected mice.

CONCLUSION

Based on these findings, isolated phage vB_PaeM_PS3 could be potentially considered for treating of P. aeruginosa infections.

摘要

背景

铜绿假单胞菌是一种医院获得性细菌,可导致多种感染。抗生素的不当使用可能导致多药耐药(MDR)铜绿假单胞菌菌株的出现。在此,分离出一种烈性噬菌体;vB_PaeM_PS3,并将其测试为替代抗生素控制铜绿假单胞菌感染的应用。

方法

使用透射电子显微镜(TEM)观察噬菌体形态。分析噬菌体的宿主范围和效价(EOP)以及噬菌体的稳定性。进行一步生长曲线以检测噬菌体的生长动力学。评估分离噬菌体对浮游细胞和生物膜的影响。对噬菌体基因组进行测序。最后,在体内确定 vB_PaeM_PS3 的治疗潜力。

结果

分离的噬菌体具有二十面体头部和可收缩的尾部,被分配到肌尾噬菌体科。噬菌体 vB_PaeM_PS3 表现出广泛的宿主范围、强大的溶菌能力和更高的环境稳定性。分离的噬菌体潜伏期短,爆发量大。重要的是,噬菌体 vB_PaeM_PS3 可有效根除细菌生物膜。vB_PaeM_PS3 的基因组由 93922 bp 的 dsDNA 组成,G+C 含量为 49.39%。它包含 171 个预测的开放阅读框(ORFs)和 14 个基因作为 tRNA。有趣的是,噬菌体 vB_PaeM_PS3 显著减弱了铜绿假单胞菌的毒力,在噬菌体治疗后,细菌感染小鼠的存活率明显提高。此外,与未治疗的感染小鼠相比,噬菌体处理的小鼠中铜绿假单胞菌的定植能力明显受损。

结论

基于这些发现,分离的噬菌体 vB_PaeM_PS3 可被认为是治疗铜绿假单胞菌感染的潜在方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1b0e/10511388/39c0a01bd593/10096_2023_4649_Fig1_HTML.jpg

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