一株铜绿假单胞菌噬菌体全基因组分析及与抗生素体外联合应用效果  

Whole Genome Analysis of a Pseudomonas aeruginosa Phage and the Effect of Combining with Antibiotics in vitro

作　　者：王晋宇 张凯川 王芮杰 高铎 蒋祺丰 贾坤[1] WANG Jinyu;ZHANG Kaichuan;WANG Ruijie;GAO Duo;JIANG Qifeng;JIA Kun(College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China)

机构地区：[1]华南农业大学兽医学院,广州510642

出　　处：《畜牧兽医学报》2024年第2期727-738,共12页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：广州市科技计划项目(SL2022E04J00201,202206010131,202201010033);广东省现代农业产业技术体系(2023KJ127)。

摘　　要：旨在为解决耐药铜绿假单胞菌感染提供不同方法。以鸡源铜绿假单胞菌为宿主菌,从医院污水中分离纯化出铜绿假单胞菌噬菌体,并进行生物学特性研究与基因组学分析,同时采用棋盘法与三种抗生素联合应用测试其效果。结果显示:从医院污水中分离到一株铜绿假单胞菌噬菌体PaVOB,TEM显示病毒颗粒具有一个平均直径为73.02 nm的二十面体头部和一个平均长度为155.57 nm的长尾部,属于有尾噬菌体目,长尾噬菌体科,具有双链DNA基因组,宿主谱较窄,耐低温(-20℃)并且在37℃达到最适温度,有良好的酸碱耐受性,在pH为3~12时保持活性,最佳MOI为0.001,一步生长曲线显示潜伏期为10 min,爆发量为50 PFU·cell^(-1),吸附性试验表明PaVOB在10 min的吸附率为99.1%。PaVOB基因组全长为72179 bp,(G+C)%含量为54.81%,与VB PaeP FBPa1亲缘关系小于0.1,ORF预测显示,PaVOB包括74个ORF,软件预测均未发现毒力基因、耐药基因和tRNA存在。此外,抗生素联合应用结果显示,PaVOB与头孢曲松联合应用有协同作用,在效价为107 PFU·mL^(-1)时与恩诺沙星产生协同作用,与庆大霉素产生无关作用。铜绿假单胞菌噬菌体PaVOB对环境适应能力强,且基因组信息清晰,能够选择性地与不同抗生素联合应用发挥协同作用,具有治疗临床耐药铜绿假单胞菌感染的可能性。This study aimed to provide different methods for solving drug-resistant Pseudomonas aeruginosa infections.P.aeruginosa phage was isolated and purified from hospital wastewater using chicken-derived P.aeruginosa as host bacteria,and biological characterization and genomic analysis were performed,while its effect was tested using a checkerboard method in combination with three antibiotics.The results showed that a strain of P.aeruginosa phage PaVOB was isolated from hospital wastewater,TEM showed that virus particles had an icosahedral head with an average diameter of 73.02 nm and a long tail with an average length of 155.57 nm,it belongs to the order Caenorhabditis elegans,family Long-tailed Phage Siphoviridae in the order Caudovirales,had a double-stranded DNA genome,had a narrow host spectrum,was resistant to low temperatures(-20℃)and reached the optimal temperature at 37℃,had good acid-base tolerance,and maintained activity at pH 3-12,the optimal MOI is 0.001,the one-step growth curve shows that the latent period is 10 min,the outbreak amount is 50 PFU·cell^(-1),and the adsorption test shows that the adsorption rate of PaVOB at 10 min is 99.1%.The full length of PaVOB genome was 72179 bp,(G+C)%content was 54.81%,with VB PaeP FBPa1 The affinity was less than 0.1.ORF prediction showed that PaVOB included 74 ORFs,and none of the software predictions revealed the presence of virulence genes,drug resistance genes and tRNAs.In addition,the results of antibiotic combination showed that PaVOB had synergistic effect with ceftriaxone,had synergistic effect with enrofloxacin at potency of 107 PFU·mL^(-1),and had unrelated effect with gentamicin.P.aeruginosa phage PaVOB is highly adaptable to the environment and has clear genomic information,which can be selectively applied in combination with different antibiotics to exert synergistic effects and has the possibility of treating clinically resistant Pseudomonas aeruginosa.

关 键 词：铜绿假单胞菌 噬菌体 生物学特性 基因组学 抗生素联合应用 

分 类 号：Q939.48[生物学—微生物学] S859.796[农业科学—临床兽医学]