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五种新型靶向沙门氏菌噬菌体的基因组和功能特征。

Genomic and functional characterization of five novel Salmonella-targeting bacteriophages.

机构信息

Department of Biotechnology and Food Microbiology, Faculty of Biotechnology and Food Sciences, Wrocław University of Environmental and Life Sciences, Wrocław, Poland.

Department of Biochemistry and Molecular Biology, Faculty of Veterinary Medicine, Wrocław University of Environmental and Life Sciences, Wrocław, Poland.

出版信息

Virol J. 2021 Sep 8;18(1):183. doi: 10.1186/s12985-021-01655-4.

Abstract

BACKGROUND

The host-unrestricted, non-typhoidal Salmonella enterica serovar Enteritidis (S. Enteritidis) and the serovar Typhimurium (S. Typhimurium) are major causative agents of food-borne gastroenteritis, and the host-restricted Salmonella enterica serovar Gallinarum (S. Gallinarum) is responsible for fowl typhoid. Increasing drug resistance in Salmonella contributes to the reduction of effective therapeutic and/or preventive options. Bacteriophages appear to be promising antibacterial tools, able to combat infectious diseases caused by a wide range of Salmonella strains belonging to both host-unrestricted and host-restricted Salmonella serovars.

METHODS

In this study, five novel lytic Salmonella phages, named UPWr_S1-5, were isolated and characterized, including host range determination by plaque formation, morphology visualization with transmission electron microscopy, and establishment of physiological parameters. Moreover, phage genomes were sequenced, annotated and analyzed, and their genomes were compared with reference Salmonella phages by use of average nucleotide identity, phylogeny, dot plot, single nucleotide variation and protein function analysis.

RESULTS

It was found that UPWr_S1-5 phages belong to the genus Jerseyvirus within the Siphoviridae family. All UPWr_S phages were found to efficiently infect various Salmonella serovars. Host range determination revealed differences in host infection profiles and exhibited ability to infect Salmonella enterica serovars such as Enteritidis, Gallinarum, Senftenberg, Stanley and Chester. The lytic life cycle of UPWr_S phages was confirmed using the mitomycin C test assay. Genomic analysis revealed that genomes of UPWr_S phages are composed of 51 core and 19 accessory genes, with 33 of all predicted genes having assigned functions. UPWr_S genome organization comparison revealed 3 kinds of genomes and mosaic structure. UPWr_S phages showed very high sequence similarity to each other, with more than 95% average nucleotide identity.

CONCLUSIONS

Five novel UPWr_S1-5 bacteriophages were isolated and characterized. They exhibit host lysis range within 5 different serovars and are efficient in lysis of both host-unrestricted and host-restricted Salmonella serovars. Therefore, because of their ability to infect various Salmonella serovars and lytic life cycle, UPWr_S1-5 phages can be considered as useful tools in biological control of salmonellosis.

摘要

背景

无宿主限制的非伤寒沙门氏菌肠炎亚种(S. Enteritidis)和伤寒亚种(S. Typhimurium)是食源性肠胃炎的主要病原体,而宿主限制的鸡白痢沙门氏菌(S. Gallinarum)则导致禽伤寒。沙门氏菌的耐药性增加导致有效治疗和/或预防选择减少。噬菌体似乎是有前途的抗菌工具,能够对抗由广泛的属于无宿主限制和宿主限制的沙门氏菌血清型的沙门氏菌菌株引起的传染病。

方法

在本研究中,分离并鉴定了五种新型裂解性沙门氏菌噬菌体,命名为 UPWr_S1-5,包括通过噬菌斑形成确定宿主范围、透射电子显微镜观察形态以及建立生理参数。此外,对噬菌体基因组进行了测序、注释和分析,并通过平均核苷酸同一性、系统发育、点图、单核苷酸变异和蛋白质功能分析将其基因组与参考沙门氏菌噬菌体进行比较。

结果

发现 UPWr_S1-5 噬菌体属于 Siphoviridae 科中的 Jerseyvirus 属。所有 UPWr_S 噬菌体均能有效地感染各种沙门氏菌血清型。宿主范围测定显示出宿主感染谱的差异,并显示出能够感染沙门氏菌肠炎亚种,如肠炎、鸡白痢、森藤伯格、斯坦利和切斯特。使用丝裂霉素 C 试验证实了 UPWr_S 噬菌体的裂解生命周期。基因组分析表明,UPWr_S 噬菌体的基因组由 51 个核心和 19 个辅助基因组成,所有预测基因中有 33 个具有指定功能。UPWr_S 基因组组织比较显示出 3 种基因组和镶嵌结构。UPWr_S 噬菌体彼此之间具有非常高的序列相似性,平均核苷酸同一性超过 95%。

结论

分离并鉴定了五种新型 UPWr_S1-5 噬菌体。它们在 5 种不同血清型中表现出宿主裂解范围,并且能够有效地裂解无宿主限制和宿主限制的沙门氏菌血清型。因此,由于它们能够感染各种沙门氏菌血清型和裂解生命周期,因此 UPWr_S1-5 噬菌体可以被视为沙门氏菌病生物防治的有用工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fccc/8425127/b106325e63fc/12985_2021_1655_Fig1_HTML.jpg

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