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噬菌体D2SVT对耐碳青霉烯类鲍曼不动杆菌具有强大的抗菌和抗生物膜活性。

Bacteriophage D2SVT exhibits potent antimicrobial and antibiofilm activity against carbapenem-resistant Acinetobacter baumannii.

作者信息

Kulshrestha Mukta, Tiwari Vishvanath

机构信息

Department of Biochemistry, Central University of Rajasthan, Ajmer 305817, India.

出版信息

J Appl Microbiol. 2025 Jun 2;136(6). doi: 10.1093/jambio/lxaf146.

Abstract

AIMS

Bacteriophages have emerged as a therapeutic option to treat multidrug-resistant (MDR) bacteria, including carbapenem-resistant Acinetobacter baumannii (CRAB). This study reports the identification and characterization of bacteriophage D2SVT, which was effective and specific against CRAB.

METHODS AND RESULTS

Thirty-five collected samples were tested for antibacterial activity against CRAB. The D2SVT phage exhibited significant lytic activity, forming larger zones and clearer plaques than the Acinetobacter phage PBAB132 in spot assays. Growth kinetics assays, CFU plating, 3,5-dinitrosalicylic acid assays, reactive oxygen species (ROS) assays, protein carbonylation assays, and membrane damage assays confirmed D2SVT's specificity to CRAB and its superior efficacy over PBAB132. Characterization using scanning electron microscopy, transmission electron microscopy, confocal microscopy, and flow cytometry revealed CRAB membrane disruption, reduced viability, and phage-cell interaction. D2SVT also exhibited anti-biofilm activity, as observed in scanning electron microscopy (SEM) analysis. D2SVT also reduced the infection of CRAB in the A549 cell line. No non-specific effects were observed on E. coli, B. subtilis, and Lactobacillus. Transmission electron microscopy (TEM) analysis classified D2SVT as a short-tailed phage with icosahedral capsid geometry (hexagonal head structure), placing it within the Caudoviricetes family. Whole genome sequencing has identified it as a phage that is closely related to phage PhiE11, and its sequence was submitted to the National Centre for Biotechnology Information (NCBI).

CONCLUSIONS

Bacteriophage D2SVT is highly specific and effective against CRAB, demonstrating strong lytic activity and anti-biofilm properties that support its potential for phage therapy.

摘要

目的

噬菌体已成为治疗多重耐药(MDR)细菌的一种治疗选择,包括耐碳青霉烯类鲍曼不动杆菌(CRAB)。本研究报告了对CRAB具有有效性和特异性的噬菌体D2SVT的鉴定和特性。

方法与结果

对收集的35个样本进行了针对CRAB的抗菌活性测试。在点滴试验中,D2SVT噬菌体表现出显著的裂解活性,形成的噬菌斑区域比不动杆菌噬菌体PBAB132更大且更清晰。生长动力学测定、CFU平板计数、3,5-二硝基水杨酸测定、活性氧(ROS)测定、蛋白质羰基化测定和膜损伤测定证实了D2SVT对CRAB的特异性及其优于PBAB132的疗效。使用扫描电子显微镜、透射电子显微镜、共聚焦显微镜和流式细胞术进行的表征揭示了CRAB膜的破坏、活力降低以及噬菌体与细胞的相互作用。如扫描电子显微镜(SEM)分析所示,D2SVT还表现出抗生物膜活性。D2SVT还降低了CRAB在A549细胞系中的感染。未观察到对大肠杆菌、枯草芽孢杆菌和乳酸杆菌的非特异性影响。透射电子显微镜(TEM)分析将D2SVT归类为具有二十面体衣壳几何结构(六角形头部结构)的短尾噬菌体,属于长尾噬菌体科。全基因组测序已将其鉴定为与噬菌体PhiE11密切相关的噬菌体,其序列已提交至美国国立生物技术信息中心(NCBI)。

结论

噬菌体D2SVT对CRAB具有高度特异性和有效性,表现出强大的裂解活性和抗生物膜特性,支持其在噬菌体治疗中的潜力。

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