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Recombination of T4-like Phages and Its Activity against Pathogenic Escherichia coli in Planktonic and Biofilm Forms

  • RESEARCH ARTICLE
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Virologica Sinica

Abstract

The increasing emergence of multi-drug resistant Escherichia coli (E. coli) has become a global concern, primarily due to the limitation of antimicrobial treatment options. Phage therapy has been considered as a promising alternative for treating infections caused by multi-drug resistant E. coli. However, the application of phages as a promising antimicrobial agent is limited by their narrow host range and specificity. In this research, a recombinant T4-like phage, named WGqlae, has been obtained by changing the receptor specificity determinant region of gene 37, using a homologous recombination platform of T4-like phages established by our laboratory previously. The engineered phage WGqlae can lyse four additional hosts, comparing to its parental phages WG01 and QL01. WGqlae showed similar characteristics, including thermo and pH stability, optimal multiplicity of infection and one-step growth curve, to the donor phage QL01. In addition, sequencing results showed that gene 37 of recombinant phage WGqlae had genetically stable even after 20 generations. In planktonic test, phage WGqlae had significant antimicrobial effects on E. coli DE192 and DE205B. The optical density at 600 nm (OD600) of E. coli in phage WGqlae treating group was significantly lower than that of the control group (P < 0.01). Besides, phage WGqlae demonstrated an obvious inhibitory effect on the biofilm formation and the clearance of mature biofilms. Our study suggested that engineered phages may be promising candidates for future phage therapy applications against pathogenic E. coli in planktonic and biofilm forms.

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References

  • Adams M (1959) Bacteriophage. Interscience Publishers, New York

    Google Scholar 

  • Bartual SG, Otero JM, Garcia-Doval C, Llamas-Saiz AL, Kahn R, Fox GC, van Raaij MJ (2010) Structure of the bacteriophage T4 long tail fiber receptor-binding tip. Proc Natl Acad Sci U S A 107:20287–20292

    Article  CAS  Google Scholar 

  • Chan BK, Abedon ST, Loc-Carrillo C (2013) Phage cocktails and the future of phage therapy. Future Microbiol 8:769–783

    Article  CAS  Google Scholar 

  • Chan BK, Turner PE, Kim S, Mojibian HR, Elefteriades JA, Narayan D (2018) Phage treatment of an aortic graft infected with Pseudomonas aeruginosa. Evol Med Public Health 2018:60–66

    Article  Google Scholar 

  • Chen M, Xu J, Yao H, Lu C, Zhang W (2016) Isolation, genome sequencing and functional analysis of two T7-like coliphages of avian pathogenic Escherichia coli. Gene 582:47–58

    Article  CAS  Google Scholar 

  • Chen M, Zhang L, Abdelgader SA, Yu L, Xu J, Yao H, Lu C, Zhang W (2017) Alterations in gp37 expand the host range of a T4-like phage. Appl Environ Microbiol. https://doi.org/10.1128/AEM.01576-17

    Article  PubMed  PubMed Central  Google Scholar 

  • Chibani-Chennoufi S, Sidoti J, Bruttin A, Kutter E, Sarker S, Brussow H (2004) in vitro and in vivo bacteriolytic activities of Escherichia coli phages: implications for phage therapy. Antimicrob Agents Chemother 48:2558–2569

    Article  CAS  Google Scholar 

  • Clark JR, March JB (2006) Bacteriophages and biotechnology: vaccines, gene therapy and antibacterials. Trends Biotechnol 24:212–218

    Article  CAS  Google Scholar 

  • Dedrick RM, Guerrero-Bustamante CA, Garlena RA, Russell DA, Ford K, Harris K, Gilmour KC, Soothill J, Jacobs-Sera D, Schooley RT, Hatfull GF, Spencer H (2019) Engineered bacteriophages for treatment of a patient with a disseminated drug-resistant Mycobacterium abscessus. Nat Med 25:730–733

    Article  CAS  Google Scholar 

  • Diarra MS, Malouin F (2014) Antibiotics in Canadian poultry productions and anticipated alternatives. Front Microbiol 5:282

    Article  Google Scholar 

  • Ebrahimi-Nik H, Bassami MR, Mohri M, Rad M, Khan MI (2018) Bacterial ghost of avian pathogenic E. coli (APEC) serotype O78:K80 as a homologous vaccine against avian colibacillosis. PLoS ONE 13:888

    Article  Google Scholar 

  • Endersen L, Buttimer C, Nevin E, Coffey A, Neve H, Oliveira H, Lavigne R, O’Mahony J (2017) Investigating the biocontrol and anti-biofilm potential of a three phage cocktail against Cronobacter sakazakii in different brands of infant formula. Int J Food Microbiol 253:1–11

    Article  CAS  Google Scholar 

  • Haq IU, Chaudhry WN, Akhtar MN, Andleeb S, Qadri I (2012) Bacteriophages and their implications on future biotechnology: a review. Virol J 9:9

    Article  Google Scholar 

  • Hashemolhosseini S, Montag D, Kramer L, Henning U (1994) Determinants of receptor specificity of coliphages of the T4 family. A chaperone alters the host range. J Mol Biol 241:524–533

    Article  CAS  Google Scholar 

  • Hermoso JA, Garcia JL, Garcia P (2007) Taking aim on bacterial pathogens: from phage therapy to enzybiotics. Curr Opin Microbiol 10:461–472

    Article  CAS  Google Scholar 

  • Hughes G, Webber MA (2017) Novel approaches to the treatment of bacterial biofilm infections. Br J Pharmacol 174:2237–2246

    Article  CAS  Google Scholar 

  • Ibrahim RA, Cryer TL, Lafi SQ, Basha EA, Good L, Tarazi YH (2019) Identification of Escherichia coli from broiler chickens in Jordan, their antimicrobial resistance, gene characterization and the associated risk factors. BMC Vet Res 15:159

    Article  Google Scholar 

  • Jenkins C (2018) Enteroaggregative Escherichia coli. Curr Top Microbiol Immunol 416:27–50 

    CAS  PubMed  Google Scholar 

  • Johnson RP, Gyles CL, Huff WE, Ojha S, Huff GR, Rath NC, Donoghue AM (2008) Bacteriophages for prophylaxis and therapy in cattle, poultry and pigs. Anim Health Res Rev 9:201–215

    Article  CAS  Google Scholar 

  • Kaistha SD, Umrao PD (2016) Bacteriophage for mitigation of multiple drug resistant biofilm forming pathogens. Recent Pat Biotechnol 10:184–194

    Article  CAS  Google Scholar 

  • Kaper JB, Nataro JP, Mobley HL (2004) Pathogenic Escherichia coli. Nat Rev Microbiol 2:123–140

    Article  CAS  Google Scholar 

  • Kellenberger E, Bolle A, Boydelatour E, Epstein RH, Franklin NC, Jerne NK, Reale Scafati A, Sechaud J (1965) Functions and properties related to the tail fibers of bacteriophage T4. Virology 26:419–440

    Article  CAS  Google Scholar 

  • Kelly D, McAuliffe O, Ross RP, Coffey A (2012) Prevention of Staphylococcus aureus biofilm formation and reduction in established biofilm density using a combination of phage K and modified derivatives. Lett Appl Microbiol 54:286–291

    Article  CAS  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  Google Scholar 

  • Li Y, Chen L, Wu X, Huo S (2015) Molecular characterization of multidrug-resistant avian pathogenic Escherichia coli isolated from septicemic broilers. Poult Sci 94:601–611

    Article  CAS  Google Scholar 

  • Liu C, Diao Y, Wang D, Chen H, Tang Y, Diao Y (2019) Duck viral infection escalated the incidence of avian pathogenic Escherichia coli in China. Transbound Emerg Dis 66:929–938

    Article  CAS  Google Scholar 

  • Lu Z, Breidt F Jr, Fleming HP, Altermann E, Klaenhammer TR (2003) Isolation and characterization of a Lactobacillus plantarum bacteriophage, phiJL-1, from a cucumber fermentation. Int J Food Microbiol 84:225–235

    Article  CAS  Google Scholar 

  • Lutful Kabir SM (2010) Avian colibacillosis and salmonellosis: a closer look at epidemiology, pathogenesis, diagnosis, control and public health concerns. Int J Environ Res Public Health 7:89–114

    Article  CAS  Google Scholar 

  • Mahichi F, Synnott AJ, Yamamichi K, Osada T, Tanji Y (2009) Site-specific recombination of T2 phage using IP008 long tail fiber genes provides a targeted method for expanding host range while retaining lytic activity. FEMS Microbiol Lett 295:211–217

    Article  CAS  Google Scholar 

  • Moulin-Schouleur M, Reperant M, Laurent S, Bree A, Mignon-Grasteau S, Germon P, Rasschaert D, Schouler C (2007) Extraintestinal pathogenic Escherichia coli strains of avian and human origin: link between phylogenetic relationships and common virulence patterns. J Clin Microbiol 45:3366–3376 

    Article  CAS  Google Scholar 

  • Nesta B, Pizza M (2018) Vaccines against Escherichia coli. Curr Top Microbiol Immunol 416:213–242

    CAS  PubMed  Google Scholar 

  • Pajunen M, Kiljunen S, Skurnik M (2000) Bacteriophage phiYeO3-12, specific for Yersinia enterocolitica serotype O:3, is related to coliphages T3 and T7. J Bacteriol 182:5114–5120

    Article  CAS  Google Scholar 

  • Rios AC, Moutinho CG, Pinto FC, Del Fiol FS, Jozala A, Chaud MV, Vila MM, Teixeira JA, Balcao VM (2016) Alternatives to overcoming bacterial resistances: state-of-the-art. Microbiol Res 191:51–80 

    Article  CAS  Google Scholar 

  • Schooley RT, Biswas B, Gill JJ, Hernandez-Morales A, Lancaster J, Lessor L, Barr JJ, Reed SL, Rohwer F, Benler S, Segall AM, Taplitz R, Smith DM, Kerr K, Kumaraswamy M, Nizet V, Lin L, McCauley MD, Strathdee SA, Benson CA, Pope RK, Leroux BM, Picel AC, Mateczun AJ, Cilwa KE, Regeimbal JM, Estrella LA, Wolfe DM, Henry MS, Quinones J, Salka S, Bishop-Lilly KA, Young R, Hamilton T (2017) Development and use of personalized bacteriophage-based therapeutic cocktails to treat a patient with a disseminated resistant Acinetobacter baumannii infection. Antimicrob Agents Chemother. https://doi.org/10.1128/AAC.00954-17

    Article  PubMed  PubMed Central  Google Scholar 

  • Stepanovic S, Vukovic D, Dakic I, Savic B, Svabic-Vlahovic M (2000) A modified microtiter-plate test for quantification of staphylococcal biofilm formation. J Microbiol Methods 40:175–179

    Article  CAS  Google Scholar 

  • Stepanovic S, Cirkovic I, Ranin L, Svabic-Vlahovic M (2004) Biofilm formation by Salmonella spp. and Listeria monocytogenes on plastic surface. Lett Appl Microbiol 38:428–432

    Article  CAS  Google Scholar 

  • Suresh G, Das RK, Kaur Brar S, Rouissi T, Avalos Ramirez A, Chorfi Y, Godbout S (2018) Alternatives to antibiotics in poultry feed: molecular perspectives. Crit Rev Microbiol 44:318–335

    Article  CAS  Google Scholar 

  • Tetart F, Repoila F, Monod C, Krisch HM (1996) Bacteriophage T4 host range is expanded by duplications of a small domain of the tail fiber adhesin. J Mol Biol 258:726–731

    Article  CAS  Google Scholar 

  • Tetart F, Desplats C, Krisch HM (1998) Genome plasticity in the distal tail fiber locus of the T-even bacteriophage: recombination between conserved motifs swaps adhesin specificity. J Mol Biol 282:543–556

    Article  CAS  Google Scholar 

  • Wais AC, Goldberg EB (1969) Growth and transformation of phage T4 in Escherichia coli B-4, Salmonella, Aerobacter, Proteus, and Serratia. Virology 39:153–161

    Article  CAS  Google Scholar 

  • Yoichi M, Abe M, Miyanaga K, Unno H, Tanji Y (2005) Alteration of tail fiber protein gp38 enables T2 phage to infect Escherichia coli O157:H7. J Biotechnol 115:101–107

    Article  CAS  Google Scholar 

  • Yu L, Wang S, Guo Z, Liu H, Sun D, Yan G, Hu D, Du C, Feng X, Han W, Gu J, Sun C, Lei L (2018) A guard-killer phage cocktail effectively lyses the host and inhibits the development of phage-resistant strains of Escherichia coli. Appl Microbiol Biotechnol 102:971–983

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This study was supported by Grants from the National Natural Science Foundation of China (U1803109), Key research and development plan of Jiangsu province (BE2019304), National Key R&D Program of China (2018YFC1602500), the Central University Basic Scientific Research Fund-Animal pathogenic bacteria (KYZ201846) and Jiangsu modern agriculture (waterfowl) industrial technology system disease prevention and control innovation team (JATS[2018]222).

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Authors and Affiliations

  1. College of Veterinary Medicine, Nan**g Agricultural University, Nan**g, 210095, China

    Min Li, Donglin Shi, Yanxiu Li, Yuyi **ao, Mianmian Chen & Wei Zhang

  2. Key Laboratory of Animal Bacteriology, Ministry of Agriculture, Nan**g, 210095, China

    Min Li, Donglin Shi, Yanxiu Li, Yuyi **ao & Wei Zhang

  3. MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nan**g Agricultural University, Nan**g, 210095, China

    Min Li, Donglin Shi, Yanxiu Li, Yuyi **ao & Wei Zhang

  4. Department of Veterinary Medicine, College of Animal Science and Technology, Zhejiang A&F University, Lin’an, Hangzhou, 311300, China

    Mianmian Chen

  5. Hackensack-Meridian Health Center for Discovery and Innovation, Nutley, NJ, 07110, USA

    Liang Chen

  6. Department of Clinical Laboratory, The Second Affiliated Hospital of Soochow University, Suzhou, 215004, China

    Hong Du

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  1. Min Li
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  2. Donglin Shi
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  7. Hong Du
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Contributions

WZ and HD conceived of the study. ML, DS, YL, YX, MC performed experiment, computational analysis. WZ and ML wrote the paper. ML and LC revised the final manuscript. All authors read and approved the final manuscript.

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Correspondence to Hong Du or Wei Zhang.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Li, M., Shi, D., Li, Y. et al. Recombination of T4-like Phages and Its Activity against Pathogenic Escherichia coli in Planktonic and Biofilm Forms. Virol. Sin. 35, 651–661 (2020). https://doi.org/10.1007/s12250-020-00233-2

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