Isolation and characterization of a novel phage targeting Klebsiella pneumoniae K2 capsular type.

IF 1.9 4区 生物学 Q3 MICROBIOLOGY
Brazilian Journal of Microbiology Pub Date : 2025-09-01 Epub Date: 2025-06-06 DOI:10.1007/s42770-025-01707-9
Rahma Kazdaghli, Donia Mourali, Lucas Mora-Quilis, Pilar Domingo-Calap, Kamel Ben-Mahrez
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引用次数: 0

Abstract

Phage therapy is a promising therapeutic weapon in the fight against multidrug-resistant Klebsiella pneumoniae infections. A Klebsiella phage, KpTRp1, was isolated from wastewater sample collected in Tunisia. It exhibited specific lytic activity against a reference strain of K. pneumoniae serotype K2. Phage isolation and enrichment were performed using the double-layer agar method. It was characterized by transmission electron microscopy, physiological investigation and genomic sequence analysis. Transmission electron micrographs of KpTRp1 showed a myovirus morphology. The optimal multiplicity of infection by KpTRp1 was 0.0001 with a latent period lasted for 30 min. Its rise phase showed a burst period of almost 150 min and released a small burst size of about six plaque-forming units per infected cell. KpTRp1 was stable over a broad pH and temperature range. The phage genome sequencing revealed a linear double-stranded DNA genome of 48,003 base pairs with a G + C content of 48.79%, defining a new species of Sircambvirus genus of Jameshumphriesvirinae subfamily. Among the 68 open reading frames identified, about half were hypothetical proteins with unknown functions, the others being involved in DNA replication and metabolism, phage structure, host recognition and lysis. Remarkably, lytic activity of KpTRp1 against K. pneumoniae serotype K2 was confirmed by genomic analysis of life cycle, making it suitable candidate for phage therapy, especially since no antibiotic resistance, or lysogenic genes were detected in the phage genome.

一种新型靶向肺炎克雷伯菌K2荚膜型噬菌体的分离与鉴定。
噬菌体治疗是对抗多药耐药肺炎克雷伯菌感染的一种很有前途的治疗武器。从突尼斯收集的废水样本中分离出克雷伯氏菌噬菌体KpTRp1。它对一株血清型K2肺炎克雷伯菌有特异的裂解活性。采用双层琼脂法分离和富集噬菌体。通过透射电镜、生理调查和基因组序列分析对其进行了鉴定。透射电镜显示KpTRp1呈肌病毒形态。KpTRp1的最佳感染倍数为0.0001,潜伏期为30 min,上升期爆发周期约为150 min,每个感染细胞释放约6个斑块形成单位的小爆发。KpTRp1在较宽的pH和温度范围内稳定。噬菌体基因组测序显示一个线性双链DNA基因组,全长48003个碱基对,G + C含量为48.79%,定义了一个新的Jameshumphriesvirinae亚科Sircambvirus属物种。在鉴定的68个开放阅读框中,大约一半是功能未知的假设蛋白,其他的涉及DNA复制和代谢、噬菌体结构、宿主识别和裂解。值得注意的是,通过生命周期基因组分析证实了KpTRp1对肺炎k血清型K2的裂解活性,使其成为噬菌体治疗的合适候选者,特别是在噬菌体基因组中未检测到抗生素耐药性或溶原基因。
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来源期刊
Brazilian Journal of Microbiology
Brazilian Journal of Microbiology 生物-微生物学
CiteScore
4.10
自引率
4.50%
发文量
216
审稿时长
1.0 months
期刊介绍: The Brazilian Journal of Microbiology is an international peer reviewed journal that covers a wide-range of research on fundamental and applied aspects of microbiology. The journal considers for publication original research articles, short communications, reviews, and letters to the editor, that may be submitted to the following sections: Biotechnology and Industrial Microbiology, Food Microbiology, Bacterial and Fungal Pathogenesis, Clinical Microbiology, Environmental Microbiology, Veterinary Microbiology, Fungal and Bacterial Physiology, Bacterial, Fungal and Virus Molecular Biology, Education in Microbiology. For more details on each section, please check out the instructions for authors. The journal is the official publication of the Brazilian Society of Microbiology and currently publishes 4 issues per year.
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