Lytic phage PF18 on its characteristics and its effect on the treatment of the systemic infection induced by Klebsiella pneumoniae in mice
ZHANG Gai1, CHEN Song-jian1, JIN Jing1, LI Zhen-jiang1, WANG Shu-wei1, HUANG De-hai1, LI Ya-hui1, 2, WANG Xiao-ting1, 2, WANG Shan-mei3, WANG Zhong-quan2
1. Department of Pathogen Biology and Immunology, Henan Medical College, Zhengzhou 451191, China; 2. Department of Pathogen Biology, Basic Medical College of Zhengzhou University, Zhengzhou 450001, China; 3. Department of Laboratory, Henan Province People?s Hospital, Zhengzhou 450001, China
Abstract:The purpose of this study is to analyze the biological characteristics of a novel isolated phage PF18 and to observe the effect of the phage on the treatment of the systemic infection induced by Klebsiella pneumoniae clinical isolate in mice. The morphology of PF18 under electron microscope and the plaque of PF18 on the lawn of K. Pneumoniae F18 were observed. The one-step growth curve and the host range of the phage were investigated. The genome of PF18 was extracted and digested with restriction enzyme. The systemic infection of mice were induced by i.p. 107 CFU of K. pneumoniae F18, 108 PFU of the phage PF18 was administered i.v. 2 hours after the infection. The survival time of the mice was observed. The phage PF18 could form clear plaques of approximately 5 mm in diameter surrounded by a wider halo on F18 lawns. PF18 belongs to the Siphoviridae family and the Caudovirales order. The phage exhibited a 17-min latent, approximately 200 PFU/cell burst size when F18 served as its host. In addition, PF18 genome can be digested by EcoRI and BamHI, but cannot be digested by HindⅢ and kpnI, so the genome of PF18 consists of dsDNA. Phage PF18 treatment can significantly improve the survival time of mice.Without treatment, all of the mice with systemic infection induced by K. pneumoniae F18. By contrast, PF18 treated mice resulted in 100% survival rate after 1 days (P<0.01) and 30% after 7 days (P<0.001). PF18, a lytic Siphoviridae phage, has a relatively short latent and a large burst size, and can be effective in the treatment systemtic infection of mice caused by K. pneumoniae clinical strain. These characteristics shall increase the posibility of using this phage as an antibacterial agent; thus, it should be further investigated.
张改, 陈松建, 靳静, 李振江, 王书伟, 黄德海, 李亚辉, 王小亭, 王山梅, 王中全. 噬菌体PF18的生物学特性及其对肺炎克雷伯菌所致小鼠全身感染疗效的初步研究[J]. 中国人兽共患病学报, 2016, 32(10): 871-875.
ZHANG Gai, CHEN Song-jian, JIN Jing, LI Zhen-jiang, WANG Shu-wei, HUANG De-hai, LI Ya-hui, WANG Xiao-ting, WANG Shan-mei, WANG Zhong-quan. Lytic phage PF18 on its characteristics and its effect on the treatment of the systemic infection induced by Klebsiella pneumoniae in mice. Chinese Journal of Zoonoses, 2016, 32(10): 871-875.
[1] Podschun R, Ullmann U. Klebsiella spp. as nosocomial pathogens: epidemiology, taxonomy, typing methods, and pathogenicity factors[J]. Clin Microbiol Rev, 1998, 11(4): 589-603. [2] Jin J, Chen S J, Zhang G, et al. Characterization of a Klebsiellalytic phage,LH-01,and a preliminary investigation of the use of this phage to treat Klebsiella pneumoniae -induced septicemia in mice[J]. J Pathog Biol, 2015, 10(1): 25-28. (in Chinese) 靳静,陈松建,张改,等. 克雷伯菌属噬菌体LH-01的生物学特性及其对败血症小鼠疗效的初步研究[J]. 中国病原生物学杂志,2015,10(1):25-28. [3] Li ZJ, Huang DH, Jin J, et al. Isolation and biological characteristics of a novel lytic phage phf168 that infects Klebsiella pneumoniae clinical isolates, and a preliminary investigation of its use to treat fatal septicemia induced by K. pneumoniae [J]. J Pathog Biol, 2016, 11(6): 495-499. (in Chinese) 李振江, 黄德海, 靳静,等. 肺炎克雷伯菌裂解性噬菌体PhF168的生物学特性及其对重症败血症小鼠疗效的初步研究[J]. 中国病原生物学杂志, 2016,11(6),495-499. [4] Zhang G, Huang DH, Jin J, et al. A preliminary study of the biological characteristics and genome of a lytic bacteriophage, LH-02, that infects Klebsiella pneumoniae [J]. J Pathog Biol, 2016, 10(1): 9-12. (in Chinese) 张改,黄德海,靳静,等. 肺炎克雷伯菌噬菌体LH-02的生物学特性及基因组初步研究[J]. 中国病原生物学杂志,2016,10(1):9-12. [5] Jin J, Lin ZJ, Wang SW, et al. Characterization of ZZ2, a lytic bacteriophage that infects clinical isolates of Acinetobacter baumannii [J]. J Pathog Biol, 2013, 8(10): 885-888. (in Chinese) 靳静, 李振江, 王书伟, 等. 鲍曼不动杆菌噬菌体ZZ2生物学特性研究[J]. 中国病原生物学杂志, 2013, 8(10): 885-888. [6] Jin J, Li ZJ, Wang SW, et al. Isolation and characterization of ZZ1, a novel lytic phage that infects Acinetobacter baumannii clinical isolates[J]. BMC Microbiol, 2012, 12: 156-163. doi:10.1186/1471-2180-12-156 [7] Bruttin A, Brüssow H. Human volunteers receiving Escherichia coli phage T4 orally: a safety test of phage therapy[J]. Antimicrob Agents Chemother, 2005, 49(7): 2874-2878. doi:10.1128/AAC.49.7.2874-2878.2005 [8] Watanabe R, Matsumoto T, Sano G, et al. Efficacy of bacteriophage therapy against gut-derived sepsis caused by Pseudomonas aeruginosa in mice[J]. Antimicrobiol Agents Chemother, 2007, 51(2): 446-452. doi:10.1128/AAC.00635-06 [9] Letkiewicz S, Miedzybrodzki R, Fortuna W, et al. Eradication of Enterococcus faecalis by phage therapy in chronic bacterial prostatitis-case report[J]. Folia Microbiol (Praha), 2009, 54(5): 457-461. doi:10.1007/s12223-009-0064-z [10] Kutateladze M, Adamia R. Bacteriophages as potential new therapeutics to replace or supplement antibiotics[J]. Trends Biotechnol, 2010, 28(12): 591-595. doi:10.1016/j.tibtech.2010.08.001 [11] Hung CH, Kuo CF, Wang CH, et al. Experimental phage therapy in treating Klebsiella pneumoniae -mediated liver abscesses and bacteremia in mice[J]. Antimicrob Agents Chemother, 2011, 55(4): 1358-1365. doi:10.1128/AAC.01123-10 [12] Zhao C, Wang L. Bacteriophage therapy, old idea, new stage[J]. Microbiol China, 2011, 38(11): 1698-1704. (in Chinese) 赵晨, 王辂. 噬菌体治疗——旧概念,新阶段[J]. 微生物学通报, 2011, 38(11):1698-1704. [13] Carlton RM. Phage therapy: past history and future prospects[J]. Arch Immunol Ther Exp, 1999, 47(5): 267-274. [14] Brussow H, Kutter E. Phage ecology[M]. In: Kutter E, Sulak-velidze A, editors. Bacteriophages: biology and applications. Boca Raton: CRC Press, 2005: 129-163.