霍氏肠杆菌噬菌体的分离鉴定及生物学特性研究

doi:10.16431/j.cnki.1671-7236.2024.01.032

Abstract

Abstract: 【Objective】 This study was aimed to explore the morphological and biological characteristics of Enterobacter hormaechei phage, and prevent and control Enterobacter hormaechei infections.【Method】 In this study, Enterobacter hormaechei was used as host bacteria.A strain of phage was isolated from sewage and silt by double-layer agar plate method and its biological characteristics were studied.The titer of the phage, temperature and pH stability, optimal multiplicity of infection (MOI) and one-step growth curve of phage were measured.【Result】 A strain of Enterobacter hormaechei phage was successfully isolated and identified and named H2.The phage formed uniform, round and transparent plaque on the double-layer agar plate.The results of transmission electron microscopy showed that the head of phage H2 was icosahedral symmetry, with a diameter of 85 nm±2 nm and a long tail with a tail filament length of 190 nm±2 nm, was a member of Caudovirales and the Mycobacteriophages family.The results of temperature and pH stability of phage H2 showed that the titer remained stable in the range of 35-55 ℃ and pH 5.0-9.0.The best MOI was 0.0001.One-step growth curve according to the best MOI was drew, and it was measured that incubation period was about 20 min, the outbreak period was about 50 min, and the lysis amount was about 68 PFU/cell.【Conclusion】 Phage H2 had strong cracking ability, thermal stability and acid-base stability, which provided a good material for the research and development of subsequent phage preparations.

Key words: phage; isolation and identification; Enterobacter hormaechei; biological characteristics

CLC Number:

YANG Tingting, FENG Zijuan, HUANG Yi, ZHOU Honghong. Isolation, Identification and Biological Characteristics of Enterobacter hormaechei Phage[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(1): 323-329.

References

[1] TOWNSEND S M, HURRELL E, CAUBILLA-BARRON J, et a1.Characterization of an extended-spectrum beta-lactamase Enterobacter hormaechei nosocomial outbreak, and other Enterobacter hormaechei misidentified as Cronobacter (Enterobacter) sakazakii[J]. Microbiology, 2008, 154(12):3659-3667.
[2] O'HARA C M, STEIGERWALT A G, HILL B C, et al.Enterobacter hormaechei, a new species of the family Enterobacteriaceae formerly known as enteric group 75[J].Journal of Clinical Microbiology, 1989, 27(9):2046-2049.
[3] SHI H, WANG K, WANG L, et al.Case report of Enterobacter hormaechei in sheep with respiratory disease and death[J].BMC Veterinary Research, 2022, 18(1):57.
[4] GOLDBERG D W, FERNANDES M R, SELLERA F B, et al.Genetic background of CTX-M-15-producing Enterobacter hormaechei ST114 and Citrobacter freundii ST265 co-infecting a free-living green turtle (Chelonia mydas)[J].Zoonoses Public Health, 2019, 66(5):540-545.
[5] ZHOU H, GUO W, XU B, et al.Screening and identification of lignin-degrading bacteria in termite gut and the construction of LiP-expressing recombinant Lactococcus lactis[J].Microbial Pathogenesis, 2017, 112:63-69.
[6] ASIMAKIS E, DOUDOUMIS V, GOUVI G, et al.Draft genome sequence of Enterobacter hormaechei ENTS, a component of the symbiotic community of tephritid flies[J].Microbiology Resource Announcements, 2019, 8(49):e01364-19.
[7] PARDON B, BUCZINSKI S.Bovine respiratory disease diagnosis:What progress has been made in infectious diagnosis?[J].Veterinary Clinics of North America.Food Animal Practice, 2020, 36(2):425-444.
[8] WANG Z, DUAN L, LIU F, et al.First report of Enterobacter hormaechei with respiratory disease in calves[J]. BMC Veterinary Research, 2020, 16(1):1.
[9] AKBARI M, BAKHSHI B, NAJAR P S.Particular distribution of Enterobacter cloacae strains isolated from urinary tract infection within clonal complexes[J].Iranian Biomedical Journal, 2016, 20(1):49-55.
[10] QIAN W, YANG M, WANG T, et al.Antibacterial mechanism of vanillic acid on physiological, morphological, and biofilm properties of carbapenem-resistant Enterobacter hormaechei[J].Journal of Food Protection, 2020, 83(4):576-583.
[11] PAAUW A, CASPERS M, LEVERSTEIN-VAN H M, et al.Identification of resistance and virulence factors in an epidemic Enterobacter hormaechei outbreak strain[J].Microbiology, 2009, 155(Pt 5):1478-1488.
[12] VANDAMME E J, MORTELMANS K.A century of bacteriophage research and applications:Impacts on biotechnology, health, ecology and the economy[J].Journal of Chemical Technology and Biotechnology, 2019, 94(2):323-342.
[13] POPESCU M, VAN B, KHOSRAVI A, et al.Bacteriophages and the immune system[J].Annual Review of Virology, 2021, 8(1):415-435.
[14] SALMOND G P, FINERAN P C.A century of the phage:Past, present and future[J].Nature Reviews Microbiology, 2015, 13(12):777-786.
[15] 吴立婷, 陆睿, 刘半红, 等.产气荚膜梭菌噬菌体及乳酸杆菌联用改善肉鸡生长性能和调节肠道菌群功能研究[J].中国畜牧兽医, 2023, 50(6):2321-2332. WU L T, LU R, LIU B H, et al.Combination of Clostridium perfringens phage and Lactobacillus improves growth performance and regulates intestinal flora function of broilers[J].China Animal Husbandry & Veterinary Medicine, 2023, 50(6):2321-2332.(in Chinese)
[16] 赵影, 刘秀侠, 李金敏, 等.一株鼠伤寒沙门氏菌噬菌体的分离鉴定及对小鼠的防制效果研究[J].中国畜牧兽医, 2018, 45(1):234-241. ZHAO Y, LIU X X, LI J M, et al.Isolation and identification of a phage against Salmonella Typhimurium and its control effect on mice[J].China Animal Husbandry & Veterinary Medicine, 2018, 45(1):234-241.(in Chinese)
[17] YANG Z Q, TAO X Y, ZHANG H, et al.Isolation and characterization of virulent phages infecting Shewanella baltica and Shewanella putrefaciens, and their application for biopreservation of chilled channel catfish (Ictalurus punctatus)[J].International Journal Food Microbiology, 2019, 292:107-117.
[18] 李陇平, 张智英.金黄色葡萄球菌烈性噬菌体的分离鉴定和最佳保存方法研究[J].中国畜牧兽医, 2011, 38(6):141-146. LI L P, ZHANG Z Y.Isolate virulent phage of Staphylococcus aureus and the study of the approach to optimal condition for preservation[J].China Animal Husbandry & Veterinary Medicine, 2011, 38(6):141-146.(in Chinese)
[19] LU Z, BREIDT F J R, FLEMING H P, et al.Isolation and characterization of a Lactobacillus plantarum bacteriophage, ΦJL-1, from a cucumber fermentation[J].International Journal of Food Microbiology, 2003, 84(2):225-235.
[20] 冯烨, 刘军, 孙洋, 等.噬菌体最新分类与命名[J].中国兽医学报, 2013, 33(12):1954-1958. FENG Y, LIU J, SUN Y, et al.The latest classification and nomenclature of bacteriophages[J].Chinese Journal of Veterinary Science, 2013, 33(12):1954-1958.(in Chinese)
[21] NTEERAWATTANAPONG N, KENGKLA K, DILOKTHORNSAKUL P, et al.Prevention and control of multidrug-resistant Gram-negative bacteria in adult intensive care units:A systematic review and network meta-analysis[J].Clinical Infectious Diseases, 2017, 64(2):51-60.
[22] GRAVEY F, CATTOIR V, ETHUIN F, et al.ramR deletion in an Enterobacter hormaechei isolate as a consequence of therapeutic failure of key antibiotics in a long-term hospitalized patient[J].Antimicrobial Agents and Chemotherapy, 2020, 64(10):962-981.
[23] YANG B, FENG Y, MCNALLY A, et al.Occurrence of Enterobacter hormaechei carrying bla_NDM-1 and bla_KPC-2 in China[J].Diagnostic Microbiology and Infectious Diseases, 2018, 90(2):139-142.
[24] 李晓宇, 李若愚, 王晓, 等.丝状噬菌体对宿主菌适应性影响的研究进展[J].中国畜牧兽医, 2023, 50(6):2496-2506. LI X Y, LI R Y, WANG X, et al.Research progress on the influence of filamentous phages on host bacterial adaptability[J].China Animal Husbandry & Veterinary Medicine, 2023, 50(6):2496-2506.(in Chinese)
[25] 王乐平, 潘艳, 谭奕舟, 等.1株大肠杆菌噬菌体vB_EcoP_GN07的分离鉴定及全基因组分析[J].中国畜牧兽医, 2023, 50(4):1522-1531. WANG L P, PAN Y, TAN Y Z, et al.Isolation, identification and genome analysis of Escherichia coli phage vB_EcoP_GN07[J].China Animal Husbandry & Veterinary Medicine, 2023, 50(4):1522-1531.(in Chinese)
[26] ROHWER F.Global phage diversity[J].Cell, 2003, 113(2):141.
[27] LIN W, LI D, GAO M, et al.Isolation, characterization and biocontrol efficacy of a T4-like phage virulent to multidrug-resistant Enterobacter hormaechei[J].Diseases of Aquat Organisms, 2021, 147:97-109.
[28] RAHIMZADEH TORABI L, DOUDI M, NAGHAVI N S, et al. Bacteriophages PφEn-CL and PφEn-HO can eliminate MDR Enterobacter cloacae and Enterobacter hormaechei isolated from burn wound infections without toxicity for human skin cells[J].FEMS Microbiology Letters, 2021, 368(20):143.
[29] 陈曹伟.霍氏肠杆菌噬菌体生物学特性及其在食品减菌中的应用研究[D].扬州:扬州大学, 2022. CHEN C W.Biological characteristics of phages against Enterobacter hormaechei and its application in food decontamination[D].Yangzhou:Yangzhou University, 2022.(in Chinese)
[30] LI H, YANG X J, ZHU X Y, et al.Isolation and characterization of broad host-range of bacteriophages infecting Cronobacter sakazakii and its biocontrol potential in dairy products[J].Quality Assurance and Safety of Cropsy & Foods, 2021, 13(3):21-44.
[31] DUC H M, SON H M, YI H P, et al.Isolation, characterization and application of a polyvalent phage capable of controlling Salmonella and Escherichia coli O157:H7 in different food matrices[J].Food Research International, 2020, 131:977-990.
[32] SHANG Y, SUN Q, CHEN H, et al.Isolation and characterization of a novel Salmonella phage vB_SalP_TR2[J].Frontiers in Microbiology, 2021, 12:1452-1462.
[33] AMARILLAS L, RUBI-RANGEL L, CHAIDEZ C, et al.Isolation and characterization of phills, a novel phage with potential biocontrol agent against multidrug-resistant Escherichia coli[J].Frontiers in Mmicrobiology, 2017, 8:1355-1372.
[34] GILL J J, HYMAN P.Phage choice, isolation, and preparation for phage therapy[J].Current Pharmaceutical Biotechnology, 2010, 11(1):2-14.

Isolation, Identification and Biological Characteristics of Enterobacter hormaechei Phage

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