科微学术

微生物学通报

食源性致病性大肠杆菌O157:H7和O55:H7特异性噬菌体的分离与鉴定
作者:
  • 申丹丹

    申丹丹

    江南大学生物工程学院 工业生物技术教育部重点实验室, 江苏 无锡 214122;江南大学粮食发酵与食品生物制造国家工程研究中心, 江苏 无锡 214122;江南大学江苏省生物活性制品加工工程技术研究中心, 江苏 无锡 214122
    在期刊界中查找
    在百度中查找
    在本站中查找
  • 兰佳鑫

    兰佳鑫

    江南大学生物工程学院 工业生物技术教育部重点实验室, 江苏 无锡 214122;江南大学粮食发酵与食品生物制造国家工程研究中心, 江苏 无锡 214122;江南大学江苏省生物活性制品加工工程技术研究中心, 江苏 无锡 214122
    在期刊界中查找
    在百度中查找
    在本站中查找
  • 李诚昊

    李诚昊

    江南大学生物工程学院 工业生物技术教育部重点实验室, 江苏 无锡 214122;江南大学粮食发酵与食品生物制造国家工程研究中心, 江苏 无锡 214122;江南大学江苏省生物活性制品加工工程技术研究中心, 江苏 无锡 214122
    在期刊界中查找
    在百度中查找
    在本站中查找
  • 陆健

    陆健

    江南大学生物工程学院 工业生物技术教育部重点实验室, 江苏 无锡 214122;江南大学粮食发酵与食品生物制造国家工程研究中心, 江苏 无锡 214122;江南大学江苏省生物活性制品加工工程技术研究中心, 江苏 无锡 214122
    在期刊界中查找
    在百度中查找
    在本站中查找
  • 李晓敏

    李晓敏

    江南大学生物工程学院 工业生物技术教育部重点实验室, 江苏 无锡 214122;江南大学粮食发酵与食品生物制造国家工程研究中心, 江苏 无锡 214122;江南大学江苏省生物活性制品加工工程技术研究中心, 江苏 无锡 214122
    在期刊界中查找
    在百度中查找
    在本站中查找
基金项目:

国家自然科学基金(31701730)


Isolation and identification of phages of food-borne pathogenic Escherichia coli O157:H7 and O55:H7
Author:
  • SHEN Dandan

    SHEN Dandan

    Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China;National Engineering Research Center for Cereal Fermentation and Food Biomanufacturing, Jiangnan University, Wuxi 214122, Jiangsu, China;Jiangsu Provincial Engineering Research Center for Bioactive Product Processing, Jiangnan University, Wuxi 214122, Jiangsu, China
    在期刊界中查找
    在百度中查找
    在本站中查找
  • LAN Jiaxin

    LAN Jiaxin

    Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China;National Engineering Research Center for Cereal Fermentation and Food Biomanufacturing, Jiangnan University, Wuxi 214122, Jiangsu, China;Jiangsu Provincial Engineering Research Center for Bioactive Product Processing, Jiangnan University, Wuxi 214122, Jiangsu, China
    在期刊界中查找
    在百度中查找
    在本站中查找
  • LI Chenghao

    LI Chenghao

    Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China;National Engineering Research Center for Cereal Fermentation and Food Biomanufacturing, Jiangnan University, Wuxi 214122, Jiangsu, China;Jiangsu Provincial Engineering Research Center for Bioactive Product Processing, Jiangnan University, Wuxi 214122, Jiangsu, China
    在期刊界中查找
    在百度中查找
    在本站中查找
  • LU Jian

    LU Jian

    Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China;National Engineering Research Center for Cereal Fermentation and Food Biomanufacturing, Jiangnan University, Wuxi 214122, Jiangsu, China;Jiangsu Provincial Engineering Research Center for Bioactive Product Processing, Jiangnan University, Wuxi 214122, Jiangsu, China
    在期刊界中查找
    在百度中查找
    在本站中查找
  • LI Xiaomin

    LI Xiaomin

    Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China;National Engineering Research Center for Cereal Fermentation and Food Biomanufacturing, Jiangnan University, Wuxi 214122, Jiangsu, China;Jiangsu Provincial Engineering Research Center for Bioactive Product Processing, Jiangnan University, Wuxi 214122, Jiangsu, China
    在期刊界中查找
    在百度中查找
    在本站中查找
  • 摘要
  • | |
  • 访问统计
  • |
  • 参考文献 [27]
  • |
  • 相似文献 [20]
  • | | |
  • 文章评论
    摘要:

    【背景】肠出血性大肠杆菌(enterohemorrhagicEscherichia coli,EHEC) O157:H7和肠致病性大肠杆菌(enteropathogenicE. coli,EPEC) O55:H7是2株常见食源性致病菌,能导致腹泻及肠道外疾病,其特异性噬菌体具有制备新型抗菌制剂的应用前景。【目的】分离能特异裂解O157:H7和O55:H7的噬菌体,并分析其生物学特性和基因组特征,探索致病性大肠杆菌防控的抗生素替代方法。【方法】利用双层平板法从环境水样中分离噬菌体,对其形态、感染复数、宿主范围、一步曲线等生物学特性进行鉴定,使用Illumina MiSeq平台对其全基因组进行测序,利用RAST、Prokka、BLASTp等软件进行生物信息学分析。【结果】分别以E. coli O157:H7和O55:H7为宿主分离出2株特异性烈性噬菌体:vB_EcoM_P251和vB_EcoM_P255,均属于肌尾病毒科(Myoviridae)。最佳感染复数均为1,在培养15 min内能以91.9%和90.8%的速率吸附到宿主细胞上,而且在37-60 ℃、pH 4.0-11.0条件下保持高且稳定的活性;P251仅对大肠杆菌O157:H7和O78:H11菌株具有感染性,P255对O55:H7、O157:H7等11株不同血清型的EHEC和EPEC均具有感染性。P251基因组全长为136 254 bp,P255基因组全长为111 068 bp,GC含量分别为37%和35%;分别含有227、173个开放阅读框(open reading frame,ORF),其中,80、73个ORF与已知功能基因具有显著相似性;P251还含有2个tRNA。比较基因组显示,P251和P255基因组在其48%的长度上共享72.24%的核苷酸同一性。【结论】分离鉴定了2株新的O157:H7和O55:H7噬菌体P251和P255,宿主范围广泛,具有防控食源性致病性大肠杆菌的潜力。

    Abstract:

    [Background] Enterohemorrhagic Escherichia coli (EHEC) O157:H7 and enteropathogenic E. coli (EPEC) O55:H7 are common food-borne pathogens, causing intestinal infection and other diseases. The specific bacteriophages are of great potential for the development of new antibacterial agents. [Objective] To isolate the phages of O157:H7 and O55:H7 and to characterize their biological and genomic characteristics for future development of phage therapy. [Methods] Phages were separated from environmental water samples by the double agar overlay plaque assay. Their biological characteristics, including morphology, multiplicity of infection (MOI), host range, as well as one-step growth curve, were studied. Their genomes were sequenced by the Illumina MiSeq and then analyzed with RAST, Prokka, BLASTp. [Results] We isolated two potent phages of Myoviridae family, vB_EcoM_P251 and vB_EcoM_P255, using E. coli O157:H7 and O55:H7 as host, respectively. Their optimal MOI were both 1, and 91.9% and 90.8% of them adsorbed to the respective host within 15 min. Their lysis activity was high and stable at 37–60 ℃ and pH 4.0–11.0. P251 was only infectious to E. coli O157:H7 and O78:H11, while P255 was infectious to 11 EHEC and EPEC strains, including O55:H7 and O157:H7. The P251 genome and P255 genome are 136 254 bp and 111 068 bp in length separately, with GC content of 37% and 35%, respectively. Their genomes contain 227 and 173 open reading frames (ORFs), separately, 80 and 73 of which share significant similarities to functional genes. Besides, P251 contains 2 tRNAs. In addition, genomes of P251 and P255 share 72.24% nucleotide identity over 48% of their length. [Conclusion] Two new O157:H7 and O55:H7 phages, P251 and P255, with strong lysis activity, wide host range, as well as a great application potential in food-borne pathogenic E. coli control, were isolated and identified.

    参考文献
    [1] Safwat Mohamed D, Farouk Ahmed E, Mohamed Mahmoud A, Abd El-Baky RM, John J. Isolation and evaluation of cocktail phages for the control of multidrug-resistant Escherichia coli serotype O104:H4 and E. coli O157:H7 isolates causing diarrhea[J]. FEMS Microbiology Letters, 2017, 365(2): fnx275
    [2] Kyle JL, Cummings CA, Parker CT, Quiñones B, Vatta P, Newton E, Huynh S, Swimley M, Degoricija L, Barker M, et al. Escherichia coli serotype O55:H7 diversity supports parallel acquisition of bacteriophage at shiga toxin phage insertion sites during evolution of the O157:H7 lineage[J]. Journal of Bacteriology, 2012, 194(8): 1885-1896
    [3] Sharma M, Ryu JH, Beuchat LR. Inactivation of Escherichia coli O157:H7 in biofilm on stainless steel by treatment with an alkaline cleaner and a bacteriophage[J]. Journal of Applied Microbiology, 2005, 99(3): 449-459
    [4] Tozzoli R, Grande L, Michelacci V, Fioravanti R, Gally D, Xu XF, La Ragione R, Anjum M, Wu GH, Caprioli A, et al. Identification and characterization of a peculiar vtx2-converting phage frequently present in verocytotoxin-producing Escherichia coli O157 isolated from human infections[J]. Infection and Immunity, 2014, 82(7): 3023-3032
    [5] Weiss-Muszkat M, Shakh D, Zhou YZ, Pinto R, Belausov E, Chapman MR, Sela S. Biofilm formation by and multicellular behavior of Escherichia coli O55:H7, an atypical enteropathogenic strain[J]. Applied and Environmental Microbiology, 2010, 76(5): 1545-1554
    [6] Zhou ZM, Li XM, Liu B, Beutin L, Xu JG, Ren Y, Feng L, Lan RT, Reeves PR, Wang L. Derivation of Escherichia coli O157:H7 from its O55:H7 precursor[J]. PLoS One, 2010, 5(1): e8700
    [7] Sulakvelidze A. Phage therapy: an attractive option for dealing with antibiotic-resistant bacterial infections[J]. Drug Discovery Today, 2005, 10(12): 807-809
    [8] Cieplak T, Soffer N, Sulakvelidze A, Nielsen DS. A bacteriophage cocktail targeting Escherichia coli reduces E. coli in simulated gut conditions, while preserving a non-targeted representative commensal normal microbiota[J]. Gut Microbes, 2018, 9(5): 391-399
    [9] Dissanayake U, Ukhanova M, Moye ZD, Sulakvelidze A, Mai V. Bacteriophages reduce pathogenic Escherichia coli counts in mice without distorting gut microbiota[J]. Frontiers in Microbiology, 2019, 10: 1984
    [10] Yi Y, Abdelhamid AG, Xu YM, Yousef AE. Characterization of broad-host lytic Salmonella phages isolated from livestock farms and application against Salmonella enteritidis in liquid whole egg[J]. LWT, 2021, 144: 111269
    [11] Sanlibaba P, TEZEL BU. Isolation of phages infecting listeria monocytogenes[J]. 2019. DOI: 10.15237/gida. GD19036
    [12] Liu M, Bischoff KM, Gill JJ, Mire-Criscione MD, Berry JD, Young R, Summer EJ. Bacteriophage application restores ethanol fermentation characteristics disrupted by Lactobacillus fermentum[J]. Biotechnology for Biofuels, 2015, 8: 132
    [13] Horiuk Y, Horiuk V, Kukhtyn M, Tsvihun A, Kernychnyi S. Characterization of lytic activity of phage SAvB14 on Staphylococcus aureus variant bovis[J]. Journal of Advanced Veterinary and Animal Research, 2020, 7(3): 509-513
    [14] 郭浩然, 许菁彦, 周洋, 李子齐, 董星星, 李锦铨. 噬菌体的分布广泛性以及口服安全性[J]. 中国食品学报, 2021, 21(2): 349-357 Guo HR, Xu JY, Zhou Y, Li ZQ, Dong XX, Li JQ. Extensive distribution of bacteriophage and the safety of oral application[J]. Journal of Chinese Institute of Food Science and Technology, 2021, 21(2): 349-357(in Chinese)
    [15] Bujak K, Decewicz P, Kaminski J, Radlinska M. Identification, characterization, and genomic analysis of novel Serratia temperate phages from a gold mine[J]. International Journal of Molecular Sciences, 2020, 21(18): 6709
    [16] Litt PK, Saha J, Jaroni D. Characterization of bacteriophages targeting non-O157 shiga toxigenic Escherichia coli[J]. Journal of Food Protection, 2018, 81(5): 785-794
    [17] Yang HJ, Liang L, Lin SX, Jia SR. Isolation and characterization of a virulent bacteriophage AB1 of Acinetobacter baumannii[J]. BMC Microbiology, 2010, 10: 131
    [18] Han H, Wei XT, Wei Y, Zhang XF, Li XM, Jiang JZ, Wang R. Isolation, characterization, and bioinformatic analyses of lytic Salmonella enteritidis phages and tests of their antibacterial activity in food[J]. Current Microbiology, 2017, 74(2): 175-183
    [19] Yıldirim Z, Sakin T, Akçelik M, Akçelik N. Identification and characterization of lytic bacteriophages specific to foodborne pathogenic Escherichia coli O157:H7[J]. Food Science and Technology International, 2021, 27(1): 56-72
    [20] Summer EJ, Gonzalez CF, Bomer M, Carlile T, Embry A, Kucherka AM, Lee J, Mebane L, Morrison WC, Mark L, et al. Divergence and mosaicism among virulent soil phages of the Burkholderia cepacia complex[J]. Journal of Bacteriology, 2006, 188(1): 255-268
    [21] Yuan YH, Gao MY, Peng Q, Wu DD, Liu PM, Wu Y. Genomic analysis of a phage and prophage from a Bacillus thuringiensis strain[J]. The Journal of General Virology, 2014, 95(Pt 3): 751-761
    [22] Lu SG, Le S, Tan YL, Zhu JM, Li M, Rao XC, Zou LY, Li S, Wang J, Jin XL, et al. Genomic and proteomic analyses of the terminally redundant genome of the Pseudomonas aeruginosa phage PaP1: establishment of genus PaP1-like phages[J]. PLoS One, 2013, 8(5): e62933
    [23] Philippe C, Levesque S, Dion MB, Tremblay DM, Horvath P, Lüth N, Cambillau C, Franz C, Neve H, Fremaux C, et al. Novel genus of phages infecting Streptococcus thermophilus: genomic and morphological characterization[J]. Applied and Environmental Microbiology, 2020, 86(13): e00227-e00220
    [24] Adriaenssens E, Brister JR. How to name and classify your phage: an informal guide[J]. Viruses, 2017, 9(4): 70
    [25] Topka G, Bloch S, Nejman-Faleńczyk B, Gąsior T, Jurczak-Kurek A, Necel A, Dydecka A, Richert M, Węgrzyn G, Węgrzyn A. Characterization of bacteriophage vB-EcoS-95, isolated from urban sewage and revealing extremely rapid lytic development[J]. Frontiers in Microbiology, 2019, 9: 3326
    [26] Zhao FY, Sun HZ, Zhou XY, Liu GQ, Li ML, Wang C, Liu S, Zhuang YT, Tong YG, Ren HY. Characterization and genome analysis of a novel bacteriophage vB_SpuP_Spp16 that infects Salmonella enterica serovar pullorum[J]. Virus Genes, 2019, 55(4): 532-540
    [27] Litt PK, Jaroni D. Isolation and physiomorphological characterization of Escherichia coli O157:H7-infecting bacteriophages recovered from beef cattle operations[J]. International Journal of Microbiology, 2017, 2017: 7013236
    引证文献
    网友评论
    网友评论
    分享到微博
    发 布
引用本文

申丹丹,兰佳鑫,李诚昊,陆健,李晓敏. 食源性致病性大肠杆菌O157:H7和O55:H7特异性噬菌体的分离与鉴定[J]. 微生物学通报, 2022, 49(8): 3358-3372

复制
分享
文章指标
  • 点击次数:290
  • 下载次数: 972
  • HTML阅读次数: 774
  • 引用次数: 0
历史
  • 收稿日期:2021-12-24
  • 录用日期:2022-03-10
  • 在线发布日期: 2022-07-28
  • 出版日期: 2022-08-20
文章二维码