Antibacterial activity evaluation of a novel K3-specific phage against <i>Acinetobacter baumannii</i> and evidence for receptor-binding domain transfer across morphologies

Xiangkuan Zheng; Meihan Liu; Pei Li; Sixiang Xu; Long Chen; Guoxin Xu; Xiaoxiao Pang; Hong Du; Yishan zheng; Xiang Huo; Zhongming Tan; Juan Li; Zhirong Li; Wei Zhang

doi:10.1016/j.virs.2024.08.002

October 2024

. doi: 10.1016/j.virs.2024.08.002

Citation: Xiangkuan Zheng, Meihan Liu, Pei Li, Sixiang Xu, Long Chen, Guoxin Xu, Xiaoxiao Pang, Hong Du, Yishan zheng, Xiang Huo, Zhongming Tan, Juan Li, Zhirong Li, Wei Zhang. Antibacterial activity evaluation of a novel K3-specific phage against Acinetobacter baumannii and evidence for receptor-binding domain transfer across morphologies .VIROLOGICA SINICA, 2024, 39(5) : 767-781. http://dx.doi.org/10.1016/j.virs.2024.08.002

一株新的K3型特异性鲍曼不动杆菌噬菌体的抗菌活性评估以及受体结合域跨形态转移的证据

郑祥宽 ^a ,
刘美含 ^a ,
李培 ^a ,
徐思翔 ^a ,
陈龙 ^b ,
徐国新 ^b ,
庞晓晓 ^b ,
杜鸿 ^c ,
郑以山 ^d ,
霍翔 ^e ,
谈忠鸣 ^e ,
李娟 ^f ,
李志荣 ^g ,
张炜 ^a,,

cstr: 32224.14.j.virs.2024.08.002

a. 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;
b. 苏州大学附属张家港医院, 临床检验科;
c. 苏州大学附属第二医院, 临床检验科;
d. 南京中医药大学附属第二医院, 重症医学科;
e. 江苏省疾病预防控制中心, 南京;
f. 中国疾病预防控制中心, 传染病预防控制所, 传染病智能追踪与预测国家重点实验室, 北京;
g 河北医科大学第二医院, 临床检验中心, 石家庄

通讯作者： 张炜, vszw@njau.edu.cn
收稿日期： 2024-04-09
录用日期： 2024-07-31

摘要

鲍曼不动杆菌（A. baumannii）因其高度耐药性而对公共卫生构成严重威胁，尤其是耐碳青霉烯类抗生素的鲍曼不动杆菌（CRAB）。在这项研究中，我们从医疗废水样本中分离出一株烈性噬菌体，命名为P1068，它能够裂解CRAB，主要针对K3型荚膜。基本特性分析显示，P1068感染鲍曼不动杆菌ZWAb014的最佳感染复数（MOI）为1，潜伏期为10分钟，并在4℃至37℃的温度范围和pH值为3-10的范围内保持稳定的活性。系统发育分析和平均核苷酸相似性分析表明，P1068可根据国际病毒分类委员会（ICTV）最新发布的病毒分类被归类为Caudoviricetes 纲Obolenskvirus属的新种。根据经典的形态学分类，P1068被鉴定为T4-like噬菌体（Myoviridae）。有趣的是，我们发现P1068的尾纤维蛋白(TFP)与T7-like噬菌体(Podoviridae) AbKT21phiIII (NC_048142.1)的TFP具有74%的覆盖度和88.99%的一致性。该发现表明，噬菌体的TFP基因可能在不同属和形态之间发生水平转移。体外抗菌试验表明，P1068对生物膜和游离态下的鲍曼不动杆菌均具有抗菌活性。在小鼠腹腔感染模型中，噬菌体保护小鼠免受鲍曼不动杆菌感染，显著降低了包括大脑、血液、肺、脾和肝脏在内的多种组织中的细菌负荷。总之，这项研究表明，噬菌体P1068是治疗碳青霉烯抗生素耐药和形成生物膜的鲍曼不动杆菌感染的候选药物，并扩展了对噬菌体TFP基因水平转移的理解。
- 噬菌体
- , 鲍曼不动杆菌
- , 碳青霉烯类抗生素耐药
- , 荚膜型
- , 尾纤维蛋白
- , 抗菌活性

Antibacterial activity evaluation of a novel K3-specific phage against Acinetobacter baumannii and evidence for receptor-binding domain transfer across morphologies

Xiangkuan Zheng ^a ,
Meihan Liu ^a ,
Pei Li ^a ,
Sixiang Xu ^a ,
Long Chen ^b ,
Guoxin Xu ^b ,
Xiaoxiao Pang ^b ,
Hong Du ^c ,
Yishan zheng ^d ,
Xiang Huo ^e ,
Zhongming Tan ^e ,
Juan Li ^f ,
Zhirong Li ^g ,
Wei Zhang ^a,,

a. College of Veterinary Medicine, Nanjing Agricultural University, Key Lab of Animal Bacteriology, Ministry of Agriculture and Rural Affairs, Nanjing, 210095, China;
b. Department of Clinical Laboratory, Zhangjiagang Hospital Affiliated to Soochow University, Zhangjiagang, 215600, China;
c. Department of Clinical Laboratory, The Second Affiliated Hospital of Soochow University, Suzhou, 215004, China;
d. Intensive Care Unit, The Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing, 210003, China;
e. Jiangsu Provincial Center for Disease Prevention and Control, Nanjing, 210009, China;
f. National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Disease, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, 102206, China;
g. Provincial Center for Clinical Laboratories, The Second Hospital of Hebei Medical University, Shijiazhuang, 050000, China

Corresponding author: Wei Zhang, vszw@njau.edu.cn
Received Date: 09 April 2024
Accepted Date: 31 July 2024

Abstract

Acinetobacter baumannii (A. baumannii) poses a serious public health challenge due to its notorious antimicrobial resistance, particularly carbapenem-resistant A. baumannii (CRAB). In this study, we isolated a virulent phage, named P1068, from medical wastewater capable of lysing CRAB, primarily targeting the K3 capsule type. Basic characterization showed that P1068 infected the A. baumannii ZWAb014 with an optimal MOI of 1, experienced a latent period of 10 min and maintained stability over a temperature range of 4-37 °C and pH range of 3-10. Phylogenetic and average nucleotide identity analyses indicate that P1068 can be classified as a novel species within the genus Obolenskvirus of the Caudoviricetes class as per the most recent virus classification released by the International Committee on Taxonomy of Viruses (ICTV). Additionally, according to classical morphological classification, P1068 is identified as a T4-like phage (Myoviridae). Interestingly, we found that the tail fiber protein (TFP) of P1068 shares 74% coverage and 88.99% identity with the TFP of a T7-like phage (Podoviridae), AbKT21phiIII (NC_048142.1). This finding suggests that the TFP gene of phages may undergo horizontal transfer across different genera and morphologies. In vitro antimicrobial assays showed that P1068 exhibited antimicrobial activity against A. baumannii in both biofilm and planktonic states. In mouse models of intraperitoneal infection, P1068 phage protected mice from A. baumannii infection and significantly reduced bacterial loads in various tissues such as the brain, blood, lung, spleen, and liver compared to controls. In conclusion, this study demonstrates that phage P1068 might be a potential candidate for the treatment of carbapenem-resistant and biofilm-forming A. baumannii infections, and expands the understanding of horizontal transfer of phage TFP genes.
- Phage
- , A. baumannii
- , Carbapenem-resistant
- , Capsule type
- , Tail fiber protein
- , Antibacterial activity

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