一株牛源变栖克雷伯菌噬菌体的分离鉴定、生物学特性及全基因组分析

doi:10.11843/j.issn.0366-6964.2025.12.038

摘要/Abstract

摘要：

旨在对1株分离出的变栖克雷伯菌噬菌体进行生物学特性及全基因组分析。使用双层平板法从四川省某奶牛场乳样中分离、纯化出1株裂解性噬菌体，测定其裂解谱、热稳定性、酸碱耐受度、最佳感染复数及一步生长曲线等生物学特性，并进行全基因组分析, 同时采用透射电镜观察噬菌体形态。结果显示，分离得到一株裂解性变栖克雷伯噬菌体vB-Kvc-Y10，噬菌斑透明且有晕环，透射电镜显示其有一丝状短尾。生物学研究结果表明，噬菌体Y10宿主特异性强，不耐高温，不耐酸，最佳感染复数为0.01。一步生长曲线结果显示：潜伏期为40 min，裂解量306。全基因组分析表明：噬菌体Y10属于自复制短尾噬菌体科，为dsDNA病毒。完整基因组大小为43 281 bp，GC含量为56.19%，51个ORFs中有29个具有可推定的功能，且具有明显的功能分区。在Y10基因组中，没有发现tRNA和毒力基因的存在。遗传进化树和ANI分析均表明，噬菌体Y10与噬菌体vB_Kpl_K59PH2(登录号：OY757063)的亲缘关系最近。综上，本研究分离的噬菌体vB-Kvc-Y10是一株高特异性的裂解性噬菌体，不具有毒力基因，具有防治细菌感染的潜力。

关键词: 变栖克雷伯菌噬菌体, 分离鉴定, 生物学特性, 全基因组分析

Abstract:

This study aimed to isolate a lytic Klebsiella variicola bacteriophage, characterize its biological properties, and conduct whole-genome analysis. The phage was isolated and purified by the double-layer agar culture method from a dairy farm in Sichuan province. Its biological characteristics, including lysis spectrum, thermal stability, acid-base tolerance, optimal infection multiplicity, and one-step growth curve, were determined. Additionally, whole-genome analysis was conducted and phage morphology was observed by transmission electron microscopy (TEM). The results showed that, the isolated lytic Klebsiella variicola bacteriophage vB_Kvc_Y10. The morphology of plaque is round and transparent with clear boundaries, and the TEM shows that Y10 has a short, filamentous tail. The biological results showed that phage Y10 has high host specificity, and is not resistant to high temperatures and acidic environment. Meanwhile, the optimal multiplicity of infection (OMOI) is 0.01. The result of one-step growth curve assay indicates that the latency period is 40 minutes, and the lysis amount is 306. Whole-genome analysis indicates that bacteriophage Y10 belongs to the family of Autographiviridaes and is a dsDNA virus. The complete genome size is 43 281 bp, with a GC content of 56.19%. Among the 51 ORFs, 29 have putative functions and exhibit distinct functional partitions. No tRNA or toxin genes are found in the Y10 genome. Both the phylogenetic tree and ANI analysis indicate that bacteriophage Y10 is closely related to bacteriophage vB_Kpl_K59PH2 (accession number: OY757063). In conclusion, bacteriophage Y10 is a highly specific lytic bacteriophage without virulence genes, presenting potential for the prevention and treatment of bacterial infections.

Key words: Klebsiella variicola bacteriophage, isolation and identification, biological properties, genome-wide analysis

中图分类号:

S852.61

曾严, 杨永根, 吴兆敏, 张洁如, 王印, 罗燕, 杨泽晓, 曹随忠, 姚学萍. 一株牛源变栖克雷伯菌噬菌体的分离鉴定、生物学特性及全基因组分析[J]. 畜牧兽医学报, 2025, 56(12): 6364-6374.

ZENG Yan, YANG Yonggen, WU Zhaomin, ZHANG Jieru, WANG Yin, LUO Yan, YANG Zexiao, CAO Suizhong, YAO Xueping. Isolation and Identification, Biological Properties, and Whole-genome Analysis of a Lytic Bovine Klebsiella variicola Bacteriophage[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(12): 6364-6374.

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编号 No.	细菌 Bacteria	宿主 Source	裂解性 Lysis
BKP	Klebsiella variicola	牛	+
KP18	Klebsiella pneumoniae	牛	-
LKP1	Klebsiella pneumoniae	林麝	-
SA1	Staphylococcus aureus	牛	-
SA2	Staphylococcus aureus	牛	-
S1	Salmonella	牛	-
S2	Salmonella	鸡	-
SC1	Shigella castellani	猪	-
A1	Acinetobacter	牛	-
BB1	Bordetella Bronchiseptica	兔	-

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