新型噬菌体mz - pa99对动物模型铜绿假单胞菌角膜炎的抗菌作用

IF 3.5 3区 医学 Q3 IMMUNOLOGY
Microbial pathogenesis Pub Date : 2025-11-01 Epub Date: 2025-07-29 DOI:10.1016/j.micpath.2025.107933
Milad Gharedaghi, Ruhollah Dorostkar, Hossein Aghamollaei, Hadi Esmaeili Gouvarchin Ghaleh, Seyed-Hashem Daryabari
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引用次数: 0

摘要

背景:铜绿假单胞菌是角膜炎的常见病原菌之一,具有耐药性。由于抗菌化合物的作用机制不同,噬菌体治疗可以成为治疗耐药感染的一种候选治疗方法。本研究的目的是分离、鉴定和研究噬菌体样本治疗铜绿假单胞菌引起的角膜炎的抗菌能力。材料与方法:采用透射电镜对伊朗德黑兰医院废水中分离的噬菌体mz - pa99进行鉴定。用斑点法证实噬菌体的存在,用双层琼脂培养法评价其滴度。通过对铜绿假单胞菌、金黄色葡萄球菌和大肠杆菌的宿主范围测定和生物膜破坏试验来评估噬菌体的裂解活性。通过全基因组测序确定噬菌体的新颖性。采用体外细胞毒性试验和兔眼刺激试验评价噬菌体的安全性。使用噬菌体滴眼液在小鼠模型中研究治疗效果,通过CFU计数量化细菌负荷,并通过组织病理学评估组织损伤。并对临床分离的假单胞菌的耐药模式进行了调查。结果:mz - pa99属斯特拉伯病毒科尾状病毒目。对mz - pa99噬菌体的全基因组分析表明,该噬菌体属于Elvirus属,具有线性双链DNA,长度为209,183 bp, GC含量为49.30%。在临床分离株上观察噬菌体的抗生物膜作用。10-1稀释(9.93 × 1011 PFU/mL)时测定噬菌体(HCE-2)的IC50。观察了环丙沙星与噬菌体滴眼液对铜绿假单胞菌性角膜炎的增效作用。结论:MIZ-Pa99对细菌性角膜炎感染有较好的控制作用。然而,抗生素和噬菌体联合治疗具有协同作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Antibacterial effect of a new phage, MIZ-Pa99, in the treatment of Pseudomonas aeruginosa keratitis in an animal model.

Background: Pseudomonas aeruginosa is one of the common bacterial agents in keratitis and is resistant to drugs. Due to the difference in the mechanism of action of antibacterial compounds, phage therapy can be a therapeutic candidate to deal with drug-resistant infections. The purpose of this study is to isolate, identify, and investigate the antimicrobial ability of a phage sample to treat keratitis caused by P. aeruginosa.

Materials and methods: The Bacteriophage (phage) MIZ-Pa99 isolated from hospital wastewater in Tehran, Iran was identified by TEM imaging. The presence of phage was confirmed by the spot technique and its titration was evaluated using the double-layer agar culture method. Phage lytic activity was evaluated through host range determination against P. aeruginosa, S. aureus, and E. coli, and biofilm destruction assays. Phage's novelty was determined through whole-genome sequencing. Phage safety was assessed using in vitro cytotoxicity assays and in vivo Draize eye irritation tests in rabbit eyes. Therapeutic efficacy was investigated in a mouse model using phage eye drops, with bacterial load quantified by CFU counts and tissue damage assessed by histopathology. Also, the antibiotic resistance pattern of Pseudomonas clinical isolates was investigated.

Results: MIZ-Pa99 belongs to the Straboviridae family and the Caudoviricetes order. Whole genome analysis of MIZ-Pa99 phage showed that it belongs to Elvirus genus, and has linear and double-stranded DNA with a length 209,183 bp and a GC content of 49.30 %. The anti-biofilm effect of phage was observed on clinical isolates. IC50 of phage (HCE-2) was determined as 10-1 dilution (9.93 × 1011 PFU/ml). The synergistic effect of ciprofloxacin and phage in the form of eye drops was observed in the improvement of keratitis caused by P. aeruginosa.

Conclusion: The results obtained in this study indicated that MIZ-Pa99 can be effective in controlling bacterial keratitis infection. However, combined antibiotic and phage treatment has a synergistic effect.

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来源期刊
Microbial pathogenesis
Microbial pathogenesis 医学-免疫学
CiteScore
7.40
自引率
2.60%
发文量
472
审稿时长
56 days
期刊介绍: Microbial Pathogenesis publishes original contributions and reviews about the molecular and cellular mechanisms of infectious diseases. It covers microbiology, host-pathogen interaction and immunology related to infectious agents, including bacteria, fungi, viruses and protozoa. It also accepts papers in the field of clinical microbiology, with the exception of case reports. Research Areas Include: -Pathogenesis -Virulence factors -Host susceptibility or resistance -Immune mechanisms -Identification, cloning and sequencing of relevant genes -Genetic studies -Viruses, prokaryotic organisms and protozoa -Microbiota -Systems biology related to infectious diseases -Targets for vaccine design (pre-clinical studies)
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