Isolation and identification of a newly discovered broad-spectrum <i>Acinetobacter baumannii</i> phage and therapeutic validation against pan-resistant <i>Acinetobacter baumannii</i>

Miaomiao Lin; Lele Xiong; Wen Li; Lingyan Xiao; Wei Zhang; Xiaogui Zhao; Yishan Zheng

doi:10.1016/j.virs.2025.06.003

August 2025

. doi: 10.1016/j.virs.2025.06.003

Citation: Miaomiao Lin, Lele Xiong, Wen Li, Lingyan Xiao, Wei Zhang, Xiaogui Zhao, Yishan Zheng. Isolation and identification of a newly discovered broad-spectrum Acinetobacter baumannii phage and therapeutic validation against pan-resistant Acinetobacter baumannii .VIROLOGICA SINICA, 2025, 40(4) : 587-600. http://dx.doi.org/10.1016/j.virs.2025.06.003

一株新发现的广谱鲍曼不动杆菌噬菌体的分离与鉴定及其对泛耐药鲍曼不动杆菌的治疗验证

蔺淼淼 ^a ,
熊乐乐 ^a ,
李文 ^b ,
肖玲燕 ^a ,
张炜 ^c ,
赵小贵 ^d ,
郑以山 ^a,,

a 南京中医药大学附属南京医院, 南京市第二医院, 重症医学科;
b 北京大学深圳医院, 重症医学科;
c 南京农业大学动物医学院, 农业农村部动物细菌学重点实验室;
d 康泰医学检验服务河北有限公司, 动物实验部

通讯作者： 郑以山, zhengyishan@njucm.edu.cn
收稿日期： 2024-10-13
录用日期： 2025-06-19

摘要

鲍曼不动杆菌（A. baumannii）因其多重/泛耐药特性构成重大临床挑战。本研究从医疗废水中分离出一株广谱裂解性鲍曼不动杆菌噬菌体P425，该噬菌体可靶向9种不同荚膜型多重耐药鲍曼不动杆菌（MDRAB）。生物学表征显示：P425在pH 3–12及4–50°C温度范围内保持活性，紫外线照射可耐受20分钟，最佳感染复数（OMOI）为0.00001。吸附动力学表明其在10分钟内吸附率>90%，一步生长曲线显示潜伏期为10分钟，裂解量为184 PFU/cell。基因组测序表明其为40,583 bp大小的双链DNA（GC含量39.39%），基因组相似性分析将其归类为Friunavirus属新种，电镜形态将其归为短尾噬菌体（Podoviridae）。值得注意的是，P425对MDRAB展现强效24小时体外抑制活性，且在MOI=0.001时与五类作用机制不同的抗生素联用呈现协同效应。安全性评估证实其无细胞毒性、溶血活性及体内外系统毒性。在小鼠感染模型中，P425可显著提高Ab25菌株（ST1791/KL101）感染小鼠的存活率，与左氧氟沙星联用可实现100%生存保护并促进免疫恢复。综上，P425作为新型裂解噬菌体，为对抗MDRAB感染提供了新策略。
- 鲍曼不动杆菌
- , 多重耐药
- , 噬菌体
- , 噬菌体疗法
- , 抗菌剂

Isolation and identification of a newly discovered broad-spectrum Acinetobacter baumannii phage and therapeutic validation against pan-resistant Acinetobacter baumannii

a. Department of Intensive Care Unit, The Second Hospital of Nanjing, Affiliated to Nanjing University of Chinese Medicine, Nanjing, 210003, China;
b. Department of Intensive Care Unit, Peking University Shenzhen Hospital, Shenzhen, 518000, China;
c. College of Veterinary Medicine, Nanjing Agricultural University, Key Lab of Animal Bacteriology, Ministry of Agriculture and Rural Affairs, Nanjing, 210095, China;
d. Department of Animal Experimentation, Kangtai Medical Laboratory Service Hebei Co., Langfang, 563000, China

Corresponding author: Yishan Zheng, zhengyishan@njucm.edu.cn
Received Date: 13 October 2024
Accepted Date: 19 June 2025

Abstract

The treatment of Acinetobacter baumannii (A. baumannii) poses significant clinical challenges due to its multidrug/pan-drug resistance. In this study, we isolated a broad-spectrum lytic A. baumannii phage, named P425, from medical wastewater, targeting nine multidrug-resistant A. baumannii (MDRAB) with diverse capsular types. Biological characterization revealed that P425 maintains activity at pH range of 3-12 and temperature range of 4-50 °C. It resists UV irradiation for 20 minutes, and had an optimal multiplicity of infection (OMOI) is 0.00001. The adsorption kinetics showed that P425 achieves > 90% within 10 minutes of incubation, and the one-step growth curve indicated a 10-min latent period, with a burst size of 184 PFU/cell. The genome sequencing results indicated that it harbors a double-stranded DNA genome of 40,583 bp with a GC content of 39.39%. Intergenomic similarity analysis classified it as a novel species within the Friunavirus genus, while electron microscopy results showed that it belongs to the Podoviridae family. Notably, P425 exhibits potent 24-h in vitro inhibitory activity against MDRAB, and demonstrates synergistic effect at an MOI of 0.001 when combined with five classes of antibiotics targeting distinct antimicrobial mechanisms. Safety evaluations confirmed the absence of cytotoxicity, hemolytic activity, or systemic toxicity both in vitro and in vivo. In mouse infection models, P425 can significantly improve the survival rates of mice infected with Ab25 (ST1791/KL101). When co-administered with levofloxacin, it achieved 100% protection against mortality and promoted immune recovery. Collectively, P425 is a prospective lytic phage that could offer novel strategies for combating MDRAB infections.
- A. baumannii
- , Multidrug resistance
- , Phage
- , Phage therapy
- , Antimicrobial agents

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