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欧文氏弧菌噬菌体phi50-12的分离与特性分析

Isolation and characterization of a Vibrio owensii phage phi50-12.

作者信息

Lin Ling-Chun, Tsai Yu-Chuan

机构信息

Masters Program in Biomedical Sciences, School of Medicine, Tzu Chi University, No. 701, Sec. 3, Zhongyang Rd., Hualien, 97004, Taiwan.

出版信息

Sci Rep. 2022 Sep 30;12(1):16390. doi: 10.1038/s41598-022-20831-2.

Abstract

Vibrio owensii is a widely distributed marine vibrio species that causes acute hepatopancreatic necrosis in the larvae of Panulirus ornatus and Penaeus vannamei, and is also associated with Montipora white syndrome in corals. We characterized V. owensii GRA50-12 as a potent pathogen using phenotypic, biochemical, and zebrafish models. A virulent phage, vB_VowP_phi50-12 (phi50-12), belonging to the N4-like Podoviridae, was isolated from the same habitat as that of V. owensii GRA50-12 and characterized. This phage possesses a unique sequence with no similar hits in the public databases and has a short latent time (30 min), a large burst size (106 PFU/infected cell), and a wide range of pH and temperature stabilities. Moreover, phi50-12 also demonstrated a strong lysis ability against V. owensii GRA50-12. SDS-PAGE revealed at least nine structural proteins, four of which were confirmed using LC-MS/MS analysis. The size of the phi50-12 genome was 68,059 bp, with 38.5% G + C content. A total of 101 ORFs were annotated, with 17 ORFs having closely related counterparts in the N4-like vibrio phage. Genomic sequencing confirmed the absence of antibiotic resistance genes or virulence factors. Comparative studies have shown that phi50-12 has a unique genomic arrangement, except for the well-conserved core regions of the N4-like phages. Phylogenetic analysis demonstrated that it belonged to a group of smaller genomes of N4-like vibrio phages. The therapeutic effect in the zebrafish model suggests that phi50-12 could be a potential candidate for application in the treatment of V. owensii infection or as a biocontrol agent. However, further research must be carried out to confirm the efficacy of phage50-12.

摘要

欧文氏弧菌是一种广泛分布的海洋弧菌,可导致 ornatus 龙虾和凡纳滨对虾幼体出现急性肝胰腺坏死,还与珊瑚的鹿角珊瑚白化综合征有关。我们使用表型、生化和斑马鱼模型将欧文氏弧菌 GRA50 - 12 鉴定为一种强效病原体。从与欧文氏弧菌 GRA50 - 12 相同的栖息地分离出一种烈性噬菌体 vB_VowP_phi50 - 12(phi50 - 12),它属于 N4 样短尾噬菌体科,并对其进行了鉴定。这种噬菌体具有独特的序列,在公共数据库中没有相似匹配,具有较短的潜伏期(30 分钟)、较大的爆发量(106 个噬菌斑形成单位/感染细胞)以及广泛的 pH 和温度稳定性。此外,phi50 - 12 对欧文氏弧菌 GRA50 - 12 也表现出很强的裂解能力。SDS - PAGE 显示至少有九种结构蛋白,其中四种通过 LC - MS/MS 分析得到确认。phi50 - 12 基因组大小为 68,059 碱基对,G + C 含量为 38.5%。总共注释了 101 个开放阅读框,其中 17 个开放阅读框在 N4 样弧菌噬菌体中有密切相关的对应物。基因组测序证实不存在抗生素抗性基因或毒力因子。比较研究表明,除了 N4 样噬菌体保守的核心区域外,phi50 - 12 具有独特的基因组排列。系统发育分析表明它属于 N4 样弧菌噬菌体中一组较小基因组的类别。斑马鱼模型中的治疗效果表明,phi50 - 12 可能是用于治疗欧文氏弧菌感染或作为生物防治剂的潜在候选物。然而,必须进行进一步的研究以确认噬菌体 phi50 - 12 的疗效。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/937b/9525291/1237f5d09385/41598_2022_20831_Fig1_HTML.jpg

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