Zhang Li-Ping, Li Chang-An, Zhao Yongda, Wang Zeqing, Wang Junjie, Song Feng-Jing, Liu Bao-Tao
College of Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China.
Qingdao Bolin Biotechnology Co., Ltd., Qingdao 266000, China.
Microorganisms. 2025 Jul 5;13(7):1587. doi: 10.3390/microorganisms13071587.
The high pathogenicity rate of carbapenem-resistant (CRPA) has resulted in substantial economic losses for humans and the breeding industry. Consequently, there is an urgent need to develop new alternatives to mitigate antibiotic use. Phage therapy has demonstrated promising results in numerous studies. In this study, lytic phages targeting CRPA were isolated from feces and river water samples in Shandong, China. A total of 94 phage strains with CRPA as hosts were obtained, exhibiting lysis rates that ranged from 29% to 76% for derived from humans and different types of animals (n = 246). We further examined five representative phages, the host bacteria of which were CRPA from clinical patients and poultry, and these phages included two myoviruses and three podoviruses. Their optimal multiplicities of infection (MOIs) ranged from 10 to 10, with latent periods of less than 5 to 15 min and burst durations of 140 to 175 min, resulting in burst sizes of 133 to 352 PFU/cell. All five phages exhibited the ability to survive at temperatures up to 60 °C and within pH levels of 3 to 11. Whole-genome sequencing revealed that these five phages were all double-stranded DNA phages and did not possess resistance genes or virulence factors. The two myoviruses, sharing similar sequences, were classified into the genus , with a size of 92,509 bp and 92,293 bp, 149 to 152 ORFs and 20 to 22 tRNAs. In contrast, the three similar podoviruses belong to the genus and all contained a perforin-lyase system, with a size of 43.35 kb, a GC content of 62%, 49 to 50 ORFs and 16 to 20 tRNAs. A spray disinfection experiment demonstrated that the phage cocktail exhibited a high sterilization effect after spraying and showed good efficacy against cement and metal surfaces. This study provides foundational information for further research into the elimination of CRPA in the environment.
耐碳青霉烯类肺炎克雷伯菌(CRPA)的高致病性给人类和养殖业造成了巨大经济损失。因此,迫切需要开发新的替代方法来减少抗生素的使用。噬菌体疗法在众多研究中已显示出有前景的结果。在本研究中,从中国山东的粪便和河水样本中分离出靶向CRPA的裂解性噬菌体。共获得94株以CRPA为宿主的噬菌体菌株,对来自人类和不同类型动物的CRPA(n = 246)的裂解率在29%至76%之间。我们进一步检测了五种代表性噬菌体,其宿主细菌为来自临床患者和家禽的CRPA,这些噬菌体包括两种肌尾噬菌体和三种短尾噬菌体。它们的最佳感染复数(MOI)范围为10至10,潜伏期小于5至15分钟,爆发持续时间为140至175分钟,导致爆发量为133至352噬菌斑形成单位/细胞。所有五种噬菌体都表现出在高达60°C的温度和pH值为3至11的范围内存活的能力。全基因组测序表明,这五种噬菌体均为双链DNA噬菌体,且不具有抗性基因或毒力因子。这两种肌尾噬菌体序列相似,被归类为 属,大小分别为92,509 bp和92,293 bp,有149至152个开放阅读框和20至22个转运RNA。相比之下,三种相似的短尾噬菌体属于 属,均含有穿孔素-裂解酶系统,大小为43.35 kb,GC含量为62%,有49至50个开放阅读框和16至20个转运RNA。喷雾消毒实验表明,噬菌体混合物喷雾后具有很高的杀菌效果,并且对水泥和金属表面显示出良好的效果。本研究为进一步研究消除环境中的CRPA提供了基础信息。