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一种具有防治对虾感染潜力的新型裂解性噬菌体Pv27的分离与鉴定

Isolation and characterization of a novel lytic bacteriophage Pv27 with biocontrol potential against infections in shrimp.

作者信息

Hien Vu Thi, Lanh Pham Thi, Pham Thao Thi Phuong, Tran Khang Nam, Duy Nguyen Dinh, Hoa Nguyen Thi, Canh Nguyen Xuan, Nguyen Quang Huy, Kim Seil, Quyen Dong Van

机构信息

Laboratory of Molecular Microbiology, Institute of Biotechnology, Vietnam Academy of Science and Technology, Hanoi, Vietnam.

University of Science and Technology of Hanoi, Vietnam Academy of Science and Technology, Hanoi, Vietnam.

出版信息

PeerJ. 2025 May 6;13:e19421. doi: 10.7717/peerj.19421. eCollection 2025.

Abstract

BACKGROUND

is a major disease-causing species of that is pathogenic to both farmed shrimp and humans. With the increasing spread of antibiotic-resistant strains, bacteriophages (or phages) are considered potential agents for biocontrol as an alternative to antibiotics. In this study, a bacteriophage capable of lysing , named Pv27, was isolated, characterized, and evaluated for its potential to control infections as a natural therapy.

METHODS

Phage Pv27 was isolated using the double-layer agar technique and its morphology was characterized by transmission electron microscopy (TEM). We further assessed the host range specificity, optimal multiplicity of infection (MOI), one-step growth kinetics, and environmental stability of Pv27 under various pH and temperature conditions. The inhibitory activity of Pv27 against was evaluated . Finally, genomic analysis of Pv27 was conducted through whole-genome sequencing, followed by functional annotation of open reading frames (ORFs) and phylogenetic analysis.

RESULTS

Phage Pv27 exhibited a Myovirus-like morphology, characterized by an icosahedral head (92.7 ± 2 nm) and a contractile tail (103 ± 11 nm), and belongs to the class Caudoviricetes. Pv27 demonstrated high lytic activity against its host cells, with a short latent period of approximately 25 minutes and a large burst size of 112 plaque-forming units (PFU) per infected cell. The phage displayed significant tolerance to a wide pH range (from 3 to 11) and remained heat-stable at temperatures up to 60 °C for 90 min. Genetically, Pv27 possesses a circular double-stranded DNA genome spanning 191,395 base pairs, with a G + C content of 35% and comprising 355 open reading frames (ORFs). Remarkably, up to 23 tRNA genes were identified in its genome, while no genes associated with antibiotic resistance, virulence, or lysogeny were detected, suggesting its potential as a valuable biocontrol agent. Results from the VIRIDIC, Basic Local Alignment Search Tool (BLAST) and phylogenetic analyses revealed that Pv27 is closely related to the two known phages, phiKT1024 and phiTY18. Several genes associated with enhanced environmental competitiveness were also identified in the Pv27 genome, including those encoding a PhoH-like phosphate starvation-inducible protein and endolysin. Phage Pv27 effectively lyses highlighting its potential as a biocontrol agent.

摘要

背景

是一种主要的致病物种,对养殖虾和人类均具有致病性。随着抗生素耐药菌株的不断传播,噬菌体被认为是一种替代抗生素的生物防治潜在媒介。在本研究中,分离出一种能够裂解的噬菌体,命名为Pv27,并对其进行了表征和评估,以确定其作为天然疗法控制感染的潜力。

方法

采用双层琼脂技术分离噬菌体Pv27,并通过透射电子显微镜(TEM)对其形态进行表征。我们进一步评估了Pv27在不同pH和温度条件下的宿主范围特异性、最佳感染复数(MOI)、一步生长动力学和环境稳定性。评估了Pv27对的抑制活性。最后,通过全基因组测序对Pv27进行基因组分析,随后对开放阅读框(ORF)进行功能注释和系统发育分析。

结果

噬菌体Pv27呈现出类似肌尾噬菌体的形态,其特征为二十面体头部(92.7±2nm)和收缩性尾部(103±11nm),属于有尾噬菌体目。Pv27对其宿主细胞表现出高裂解活性,潜伏期约为25分钟,每个感染细胞的爆发量很大,为112个噬菌斑形成单位(PFU)。该噬菌体在较宽的pH范围内(从3到11)表现出显著的耐受性,在高达60°C的温度下90分钟内保持热稳定性。从基因上看,Pv27拥有一个跨度为191,395个碱基对的环状双链DNA基因组,G + C含量为35%,包含355个开放阅读框(ORF)。值得注意的是,在其基因组中鉴定出多达23个tRNA基因,同时未检测到与抗生素耐药性、毒力或溶原性相关的基因,表明其作为一种有价值的生物防治剂的潜力。VIRIDIC、基本局部比对搜索工具(BLAST)和系统发育分析的结果表明,Pv27与两种已知的噬菌体phiKT1024和phiTY18密切相关。在Pv27基因组中还鉴定出几个与增强环境竞争力相关的基因,包括编码类PhoH磷酸盐饥饿诱导蛋白和内溶素的基因。噬菌体Pv27有效地裂解了,突出了其作为生物防治剂的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2cda/12063606/2fd93b6f3af9/peerj-13-19421-g001.jpg

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