宽谱鸭疫里默氏菌噬菌体筛选及生物学特性研究

doi:10.16431/j.cnki.1671-7236.2023.03.031

摘要/Abstract

摘要： 【目的】筛选宽谱鸭疫里默氏菌噬菌体,作为治疗用噬菌体组合的候选毒株。【方法】以不同含量的噬菌体进行噬斑分析测定35株噬菌体的噬菌谱;利用透射电镜观察、热与酸碱稳定性评估、最佳感染复数(MOI)测定、一步生长试验、杀菌试验分析噬菌体的形态结构和部分生物学特性。【结果】35株噬菌体中vB_RanS_202的裂解谱最宽,能裂解42%(42/100)的鸭疫里默氏菌,包括血清1、2、6、10、11和13型共6种血清型菌株。该噬菌体由直径约60 nm的截面为六边形的头部和长约220 nm的尾部组成,不耐热,效价为1.0×10⁷ PFU/mL的噬菌体,经50 ℃水浴80 min或60 ℃水浴20 min,即降为1.8×10⁵~2.6×10⁵ PFU/mL。在pH 5.0~9.0的环境中能维持活性。以RAf488为宿主菌测得其最佳MOI为0.01。一步生长曲线显示,其潜伏期和暴发期分别为60和75 min,平均裂解量约为150 PFU/cell。在宿主菌RAf488中的杀菌试验结果显示,MOI在0.004~0.016时,vB_RanS_202均具有杀菌作用。【结论】噬菌体vB_RanS_202具有宽裂解谱的特点,具备在液体培养基中的杀菌能力。本研究结果为后续噬菌体治疗用毒株的筛选提供参考。

关键词: 鸭疫里默氏菌; 噬菌体; 噬菌谱; 生物学特性

Abstract: 【Objective】 This study was aimed to screen some broad-spectrum Riemerella anatipestifer phages as candidates for therapeutic phage combinations.【Method】 The host range of 35 phages were determined by spot tests with different contents of phages.The methods of transmission electron microscope observation,thermal sensitivity and pH stability evaluation,determination of the best multiplicity of infection (MOI) in the plural,one-step growth curve drawing and sterilization experiment were used to analyze the morphological features and some biological characteristics of phage.【Result】 Phage vB_RanS_202 had the broadest cleavage spectrum among 35 phages,which could cleave 42% (42/100) of Riemerella anatipestifer strains including 6 serotypes of 1,2,6,10,11 and 13.The phage had a hexagonal head with approximately 60 nm in diameter and a long tail with 220 nm in length,which was not heat-resistant.The titer of phage decreased from 1.0×10⁷ PFU/mL to 1.8×10⁵-2.6×10⁵ PFU/mL after 80 min of 50 ℃ water bath or 20 min of 60 ℃.The phage could maintain its activity at pH 5.0 to pH 9.0.Using RAf488 as host bacteria,the optimal MOI of the phage was 0.01.One-step growth curve indicated that the latent period of phage was 60 min,the outbreak period was 75 min,and the outbreak quantity was 150 PFU/cell.The results of bactericidal experiment against RAf488 showed that phage had good germicidal efficacy when MOI was 0.004 to 0.016.【Conclusion】 Phage vB_RanS_202 had a wide lysis spectrum and the ability of bactericidal in liquid.The results of this study provided a reference for the subsequent screening of phage therapeutic strains.

Key words: Riemerella anatipestifer; phage; phagocytosis spectrum; biological characteristics

中图分类号:

Q939.48

程龙飞, 廖维连, 陈红梅, 江南松, 傅秋玲, 万春和, 刘荣昌, 施少华, 黄瑜, 傅光华. 宽谱鸭疫里默氏菌噬菌体筛选及生物学特性研究[J]. 中国畜牧兽医, 2023, 50(3): 1169-1176.

CHENG Longfei, LIAO Weilian, CHEN Hongmei, JIANG Nansong, FU Qiuling, WAN Chunhe, LIU Rongchang, SHI Shaohua, HUANG Yu, FU Guanghua. Screening and Biological Characteristics of Broad-spectrum Riemerella anatipestifer Phage[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1169-1176.

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