沙门氏菌高效价宽谱噬菌体的分离纯化及生物学特性测定

doi:10.16431/j.cnki.1671-7236.2021.07.034

摘要/Abstract

摘要： 为探索控制沙门氏菌感染的新途径，本试验进行了高效价宽裂解谱噬菌体的分离。首先利用点斑试验和成斑率（EOP）测定法对环境样品进行了沙门氏菌噬菌体的分离纯化及裂解效价和裂解谱的测定，然后对其中1株效价较高、裂解谱较广的噬菌体进行了电镜形态特征观察、最佳感染复数测定、一步生长曲线测定及温度、pH和紫外线敏感性的测定。结果显示，该株噬菌体对不同沙门氏菌的裂解率不同，裂解效价在10⁹~10¹² PFU/mL；对检测的32株沙门氏菌菌株的裂解率为59.4%；透射电镜下观察到该噬菌体为头部直径约70 nm、尾部长约140 nm的长尾噬菌体；以鼠伤寒沙门氏菌ATCC 14028为宿主菌测得最佳感染复数为0.01；在宿主菌鼠伤寒沙门氏菌ATCC 14028中的一步生长曲线显示，潜伏期为20 min，暴发期为100 min，暴发量为113 PFU/cell；在40~60 ℃效价稳定在10¹⁰ PFU/mL，pH 5.0~13.0效价在10⁶ PFU/mL以上，紫外线照射50 min效价仍有10⁶ PFU/mL。综上可知，该株噬菌体分离株具有效价高、裂解谱宽且对温度、pH、紫外线稳定性良好等特性，符合噬菌体制剂的相关特征，具备作为噬菌体制剂后备菌株开发的潜力。

关键词: 沙门氏菌; 噬菌体; 分离纯化; 生物学特性

Abstract: In order to explore a new way to control Salmonella infection,phages with high titer and wide lysis spectrum were isolated in this experiment.Firstly,spot test and efficiency of plating (EOP) test were used to isolate and purify Salmonella phage and determine its lysis titer and lysis spectrum.Then the morphological characteristics of one of the phages with high titer and wide lysis spectrum was further observed by electron microscope,and detected the multiplicity of infection(MOI),one-step growth curve,along with temperature,pH and UV sensitivity.The results showed that the lysis rate of the bacteriophage to different host bacteria was different with lysis titer from 10⁹ to 10¹² PFU/mL.The lysis rate of 32 Salmonella strains detected was 59.4%.It was observed that the phage had a long tail phage with a head diameter of about 70 nm and a tail length of about 140 nm under transmission electron microscope.The best MOI was 0.01 when measured by host of Salmonella Typhimurium ATCC 14028,and one-step growth curve showed that the latent period was 20 min,the burst period was 100 min,and the burst size was 113 PFU/cell.The titer was stable at 10¹⁰ PFU/mL at 40-60 ℃,the titer was above 10⁶ PFU/mL at pH 5.0-13.0,and the titer was still 10⁶ PFU/mL after UV irradiation for 50 min.Therefore,the phage had the characteristics of high effective lysis,wide lysis spectrum,good stability to temperature,pH,and UV,which was in line with the relevant characteristics of phage preparations,and had the potential to be developed as a backup strain for phage preparations.

Key words: Salmonella; phage; isolation and purification; biological characteristics

中图分类号:

S852.6

唐宁, 殷茵, 任慧英, 张灿, 刘文华. 沙门氏菌高效价宽谱噬菌体的分离纯化及生物学特性测定[J]. 中国畜牧兽医, 2021, 48(7): 2577-2583.

TANG Ning, YIN Yin, REN Huiying, ZHANG Can, LIU Wenhua. Isolation,Purification and Detection of Biological Characteristics of a High-titer and Wide-spectrum Salmonella Phage[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(7): 2577-2583.

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