一株广谱沙门菌噬菌体的分离鉴定及其生物学特性

doi:10.11843/j.issn.0366-6964.2024.09.030

摘要/Abstract

摘要：

噬菌体能够特异性裂解靶向细菌, 被认为是抗生素有效替代者。为了筛选到有效裂解沙门菌的噬菌体, 为探索新的沙门菌防控方法提供理论依据和参考。本研究用丝裂霉素C诱导并分离纯化沙门菌噬菌体, 通过噬菌斑与电子显微镜观察、宿主谱测定、生物学特性测定以及对该噬菌体的全基因组进行测序分析评估该噬菌体。结果显示: 成功诱导出1株广谱的长尾沙门菌噬菌体, 命名为SP18-108, 该噬菌体能够裂解至少14种不同血清型沙门菌; 在30~40 ℃及pH 4~12条件下活性稳定; 全基因组分析结果显示, 该噬菌体全长为43 250 bp, GC%含量为49.96%, 已知功能编码序列(coding sequences, CDS)占比为52%。系统发育进化树分析表明, 该噬菌体属于长尾病毒科(Siphoviridae)。本研究诱导过程中分离出1株不具有溶源基因的广谱沙门菌噬菌体, 为进一步研究沙门菌抗菌剂提供了良好的材料。

关键词: 沙门菌, 噬菌体, 生物学特性, 全基因组分析

Abstract:

Bacteriophages can specifically lyse target bacteria and are usually considered as effective alternatives to antibiotics. In order to screen for bacteriophages that are efficient to lyse Salmonella, and provide experimental data and reference for exploring the prevention and control method of Salmonella. In this study, Salmonella phage was induced and purified by mitomycin C. Phage was evaluated by plaque and electron microscopy observation, host spectrum determination, biological characterization and whole genome sequencing analysis. A broad-spectrum Salmonella long-tail phage was successfully induced, named SP18-108, which could lyse at least 14 different serotypes of Salmonella. The activity is stable at 30-40℃ and pH 4-12.The whole genome analysis showed that the length of the phage was 43 250 bp, and the GC content was 49.96%, the proportion of known coding sequences (CDS) was 52%. Phylogenetic tree analysis showed that phage SP18-108 belonged to the (Siphoviridae). In summary, in this study, we isolated a broad-spectrum Salmonella bacteriophage without lysogenic genes, which providing a good material for further research on Salmonella antibacterial agents.

Key words: Salmonella, phage, biological characteristics, whole genome analysis

中图分类号:

Q939.48

刘炜, 马嘉怡, 耿浩宇, 谢添, 苗苏南, 廖宗杰, 耿士忠. 一株广谱沙门菌噬菌体的分离鉴定及其生物学特性[J]. 畜牧兽医学报, 2024, 55(9): 4061-4068.

Wei LIU, Jiayi MA, Haoyu GENG, Tian XIE, Sunan MIAO, Zongjie LIAO, Shizhong GENG. Isolation, Identification and Characterization of a Broad Spectrum Salmonella Phage[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(9): 4061-4068.

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沙门菌Salmonella			噬菌体Bacteriophage
血清型Serotype	表面抗原Surface antigen	数量Amount	SP18-108		SP13
血清型Serotype	表面抗原Surface antigen	数量Amount	裂解能力Lysis ability	裂解率/% Lysis rate	裂解能力Lysis ability	裂解率/% Lysis rate
鼠伤寒沙门菌Salmonella Typhimurium	1, 4, 5, 12	37	+++ 37	100(37/37)	+++ 36	97.30(36/37)
肠炎沙门菌Salmonella Enteritidis	1, 9, 12	34	+++ 34	100(34/34)	+++ 27	79.41(27/34)
鸡白痢沙门菌Salmonella Pullorum	1, 9, 12	30	+++ 30	100(30/30)	+++ 25	83.33(25/30)
印第安纳沙门菌Salmonella Indiana	1, 4, 12	26	+++ 21	80.77(21/26)	+++ 18	69.2(18/26)
都柏林沙门菌Salmonella Dublin	1, 9, 12	23	+++ 18	78.26(18/23)	++ 14	60.86(14/23)
伤寒沙门菌Salmonella Typhi	9, 12, Vi	20	++ 15	75.00(15/20)	-	-
甲副伤寒沙门菌Salmonella Paratyphi a	1, 2, 12	16	++ 10	62.50(10/16)	-	-
乙副伤寒沙门菌Salmonella Paratyphi b	1, 4, 5, 12	11	++ 7	63.64(7/11)	-	-
德尔卑沙门菌Salmonella derby	1, 4, 12	10	++ 6	60.00(6/10)	-	-
肯塔基沙门菌Salmonella Kentucky	1, 6, 12	7	++ 3	42.86(3/7)	-	-
海德尔堡沙门菌Salmonella Heidelberg	4, 5, 12	5	++ 2	40.00(2/5)	-	-
鸭沙门菌Salmonella Anatum	3, 10	4	+ 1	25.00(1/4)	-	-
伦敦沙门菌Salmonella Iondon	3, 10	10	+ 2	20.00(2/10)	-	-
火鸡沙门菌Salmonella meleagridis	3, 10	10	+ 1	10.00(1/10)	-	-
里森沙门菌Salmonella Rissen	6, 7	3	0	-	-	-

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