1株沙门氏菌噬菌体的分离鉴定和特性分析

doi:10.16431/j.cnki.1671-7236.2023.10.031

Abstract

Abstract: 【Objective】 It was aimed to isolate a phage with effective cracking for Salmonella,and provide experimental data and reference for exploring the countermeasure method of Salmonella.【Method】 Salmonella Pullorum X-1014 was used as the host bacteria,and the phage was isolated and purified from sewage by double-layer plate method.The phage was evaluated by plaque and electron microscopy,host spectrum,biological characteristics test,antibacterial test in vitro and whole genome sequencing analysis.【Result】 A lytic phage was isolated and purified,named ZH5 (GenBank accession No.:OM864357.1),which could lyse Escherichia coli and Salmonella.The results of double-layer plate method showed that the plaque was transparent and the edge was clear.Electron microscopy showed that phage ZH5 had an icosahedral head with a straight diameter of about 50 nm,a tail with a length of about 20 nm.The phage ZH5 could lyse three strains of Salmonella and one strain of Escherichia coli.The results of biological characteristics showed that the optimal multiplicity of infection was 10,the incubation period was 20 min and the outbreak period was 60 min,and the lysis volume was about 139 PFU/cell.The acid-base tolerance range was pH 4.0-10.0,and it could survive for 30 min in the environment of 4-50 ℃.Phage ZH5 showed the best inhibitory effect on Salmonella X-1014 and CVCC 1806 when the multiplicity of infection was 10.The whole genome analysis showed that the length of the phage was 42 949 bp,and the GC content was 51.41%,of which the proportion of known functional coding sequences (CDS) was 52%.Phage linear alignment showed that the genome of phage ZH5 was the most similar to the Escherichia phage Minorna(identity 94.19%;cover 84%).Phylogenetic tree analysis showed that phage ZH5 belonged to the Drulisvirus genus.【Conclusion】 A phage capable of cross-host lysis of Salmonella and Escherichia coli was isolated,which provided a good theoretical material for further research on Salmonella antibacterial agents.

Key words: Salmonella; phage; biological characteristics; in vitroantibacterial effect; whole-genome analysis

CLC Number:

S852.61

LIAO Binru, YU Chunyan, HUANG Qiaochen, MO Yupeng, HUANG Weiqian, LI Lei, HAN Kaiou, WANG Leping, LI Xun, WANG Xiaoye. Isolation,Identification and Characterization of a Strain of Salmonella Phage[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(10): 4168-4178.

References

[1] 陈伟,赵学峰,徐鸿,等.山东省部分规模化鸡场鸡白痢血清学调查[J].家禽科学,2021,45(10):38-41. CHEN W,ZHAO X F,XU H,et al.Serological investigation of pullorum disease in some large-scale chicken farms in Shandong provice[J].China Poultry Science,2021,10:38-41.(in Chinese)
[2] ISAEV A B,MUSHAROVA O S,SEVERIONV K V.Microbial arsenal of antiviral defenses——Part Ⅰ[J].Biochemistry (Moscow),2021,86(3):319-337.
[3] 陶程琳,王少辉,张耀东,等.噬菌体疗法预防细菌性感染的研究进展及发展方向[J].中国兽医科学,2020,50(9):1167-1175. TAO C L,WANG S H,ZHANG Y D,et al.Advances and development direction of bacteriophage therapy for preventing bacterial infection[J].Chinese Veterinary Science,2020,50(9):1167-1175.(in Chinese)
[4] ALISK J,ICZKOWSKI K,RAPOPORT A,et al.Bacteriophages show promise as antimicrobial agents[J]. Journal of Infection,1998,36(1):5-15.
[5] ATTERBURY R J,VAN BERGEN M,ORITIZ F,et al.Bacteriophage therapy to reduce Salmonella colonization of broiler chickens[J].Applied and Environmental Microbiology,2007,73(14):4543-4549.
[6] DUC H M,SON H M,HONJOH K,et al.Isolation and application of bacteriophages to reduce Salmonella contamination in raw chicken meat[J].LWT-Food Science and Technology,2018,91(1):353-360.
[7] LIM T H,KIM M S,LEE D H,et al.Use of bacteriophage for biological control of Salmonella Enteritidis infection in chicken[J].Research in Veterinary Science,2012,93(3):1173-1178.
[8] KAZIBWE G,KATAMI P,ALTNAITWE R,et al.Bacteriophage activity against and characterisation of avian pathogenic Escherichia coli isolated from colibacillosis cases in Uganda[J].PLoS One,2020,15(12):e0239107.
[9] 韩传银,张飞雄,童贻刚.噬菌体浓缩方法的比较[J].生物技术通讯,2013,24(5):695-697. HAN C Y,ZHANG F X,TONG Y G.Comparison of different bacteriophage concentration methods[J].Letters in Biotechnology,2013,24(5):695-697.(in Chinese)
[10] ZHANG C,YUAN J,GUO C,et al.Identification and complete genome of lytic "Kp34likevirus" phage vB_KpnP_Bp5 and therapeutic potency in the treatment of lethal Klebsiella pneumoniae infections in mice[J].Virus Research,2021,297(198348):1-11.
[11] 韦德源,陆泽宁,张聪,等.产志贺毒素多重耐药大肠杆菌噬菌体的生物学特性及全基因组分析[J].中国畜牧兽医,2020,47(2):352-362. WEI D Y,LU Z N,ZHANG C,et al.Biological characteristics and genome analysis of Shiga toxin-producing and multidrug-resistant Escherichia coli phages[J].China Animal Husbandry & Veterinary Medicine,2020,47(2):352-362.(in Chinese)
[12] 程龙飞,廖维连,陈红梅,等.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 L F,LIAO W L,CHEN H M,et al.Screening and biological characteristics of broad-spectrum Riemerella anatipestifer phage[J].China Animal Husbandry & Veterinary Medicine,2023,50(3):1169-1176.(in Chinese)
[13] 赵玉,党瑞莹,王紫阳,等.一株裂解性鸡白痢沙门氏菌噬菌体的分离及生物学特性分析[J].黑龙江畜牧兽医,2023,2:82-85. ZHAO Y,DANG R Y,WANG Z Y,et al.Isolation and biological characterization of a strain of lytic phage of Salmonella enterica subsp.enterica serovar Pullorum[J].Heilongjiang Animal Science and Veterinary Medicine,2023,2:82-85.(in Chinese)
[14] 赵远洋,舒梅,吴国平,等.沙门氏菌烈性噬菌体JN-S202001的分离、鉴定及其应用于即食鸭脖的抑菌效果[J].江西农业大学学报,2022,44(6):1520-1528. ZHAO Y Y,SHU M,WU G P,et al.A virulent bacteriophage against Salmonella:Isolation,characterization and application as a biocontrol agent in ready-to-eat duck neck[J].Acta Agriculturae Universitatis Jiangxiensis,2022,44(6):1520-1528.(in Chinese)
[15] 包红朵,张鹏禹,周艳,等.沙门氏菌裂解性噬菌体的分离鉴定及其生物学特性[J].食品科学,2015,36(5):131-136. BAO H D,ZHANG P Y,ZHOU Y,et al.Isolation and biological characteristics of lytic Salmonella bacteriophage[J].Food Science,2015,36(5):131-136.(in Chinese)
[16] 周祥莹,张灿,刘文华,等.鸡白痢沙门氏菌及其噬菌体vB-SpuS-Spp4的分离鉴定[J].病毒学报,2018,34(2):239-244. ZHOU X Y,ZHANG C,LIU W H,et al.Isolation and identification of Salmonella Pullorum and its bacteriophage vB-SpuS-Spp4[J].Chinese Journal of Virology,2018,34(2):239-244.(in Chinese)
[17] 汪祥燕,辛国芹,董佩佩,等.1株宽噬菌谱鸡白痢沙门氏菌噬菌体的分离及生物学特性分析[J].中国畜牧兽医,2018,45(9):2575-2581. WANG X Y,XIN G Q,DONG P P,et al.Isolation and biological properties analysis of a Salmonella Pullorum lytic phage with wide host range[J]. China Animal Husbandry & Veterinary Medicine,2018,45(9):2575-2581.(in Chinese)
[18] 周雨晴,马东鑫,吴洁,等.宽谱大肠杆菌O157:H7 CVCC4050噬菌体Vb_EcoM_GXBP08的分离鉴定及特性分析[J].中国畜牧兽医,2022,49(7):2735-2745. ZHOU Y Q,MA D X,WU J,et al.Isolation,identification and characterization of broad-spectrum Escherichia coli O157:H7 CVCC4050 phage vB_EcoM_GXBP08[J].China Animal Husbandry & Veterinary Medicine,2022,49(7):2735-2745.(in Chinese)
[19] 赵鹏昊.ETEC K88噬菌体PE-1的分离表征及体外抑菌效果评价[D].哈尔滨:东北农业大学,2019. ZHAO P H.Isolation,characterization and in vitro antibacterial activity evaluation of ETEC K88 phage PE-1[D].Harbin:Northeast Agricultural University,2019.(in Chinese)
[20] 王心舞.噬菌体鸡尾酒-肠道菌群移植对细菌性结肠炎实验治疗及作用的分子机制[D].长春:吉林大学,2022. WANG X W.Phage cocktail-intestinal microbiota transplantation for experimental treatment of bacterial colitis and its molecular mechanism[D].Changchun:Jilin University,2022.(in Chinese)
[21] 张慧君.噬菌体S528及其诱导猪霍乱沙门氏菌产生噬菌体抗性机制的初步研究[D].大连:大连理工大学,2022. ZHANG H J.Salmonella Choleraesuis phage S528 and preliminaray study on the mechanism of phage resistance[D].Dalian:Dalian University of Technology,2022.(in Chinese)
[22] 王玉晨.嗜盐古生菌温和病毒SNJ1复制区的鉴定、应用及超感染免疫性的相关研究[D].武汉:武汉大学,2016. WANG Y C.Identification and application of the replicon region of haloarchaeal temperate virus SNJ1 and the study on its superinfection exclusion[D].Wuhan:Wuhan University,2016.(in Chinese)
[23] 聂若男.沙门氏菌噬菌体的分离鉴定、性质分析及其在食品控制中的应用[D].武汉:华中农业大学,2019. NIE R N.Isolation,identification and characterization of Salmonella phage and its application in food control[D].Wuhan:Huazhong Agricultural University,2019.(in Chinese)
[24] 伍亚云,黄勋.噬菌体治疗细菌感染的研究进展[J].中国感染控制杂志,2021,20(2):186-190. WU Y Y,HUANG X.Advances in the treatment of bacterial infections by phages[J].Chinese Journal of Infection Control,2021,20(2):186-190.(in Chinese)
[25] LU M J,STIERHOF Y D,HENNING U.Location and unusual membrane topology of the immunity protein of the Escherichia coli phage T4[J].Journal of Virology,1993,67(8):4905-4913.
[26] MACIEJEWSKA B,OLSZAK T,DRULIS-KAWA Z.Applications of bacteriophages versus phage enzymes to combat and cure bacterial infections:An ambitious and also a realistic application?[J].Applied Microbiology and Biotechnology,2018,102(6):2563-2581.

Isolation,Identification and Characterization of a Strain of Salmonella Phage

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