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Lysogenic conversion of atypical enteropathogenic Escherichia coli (aEPEC) from human, murine, and bovine origin with bacteriophage Φ3538 Δstx2::cat proves their enterohemorrhagic E. coli (EHEC) progeny.
Int J Med Microbiol. 2018 Oct; 308(7):890-898.IJ

Abstract

Bacteriophages play an important role in the evolution of bacterial pathogens. A phage-mediated transfer of stx-genes to atypical enteropathogenic E. coli (aEPEC) which are prevalent in different hosts, would convert them to enterohemorrhagic E. coli (EHEC). We decided to confirm this hypothesis experimentally to provide conclusive evidence that aEPEC isolated from different mammalian hosts are indeed progenitors of typical EHEC which gain the ability to produce Shiga-Toxin by lysogeny with stx-converting bacteriophages, utilizing the model phage Φ3538 Δstx2::cat. We applied a modified in vitro plaque-assay, using a high titer of a bacteriophage carrying a deletion in the stx2 gene (Φ3538 Δstx2::cat) to increase the detection of lysogenic conversion events. Three wild-type aEPEC strains were chosen as acceptor strains: the murine aEPEC-strain IMT14505 (sequence type (ST)28, serotype Ont:H6), isolated from a striped field mouse (Apodemus agrarius) in the surrounding of a cattle shed, and the human aEPEC-strain 910#00 (ST28, Ont:H6). The close genomic relationship of both strains implies a high zoonotic potential. A third strain, the bovine aEPEC IMT19981, was of serotype O26:H11 and ST21 (STC29). All three aEPEC were successfully lysogenized with phage Φ3538 Δstx2::cat. Integration of the bacteriophage DNA into the aEPEC host genomes was confirmed by amplification of chloramphenicol transferase (cat) marker gene and by Southern-Blot hybridization. Analysis of the whole genome sequence of each of the three lysogens showed that the bacteriophage was integrated into the known tRNA integration site argW, which is highly variable among E. coli. In conclusion, the successful lysogenic conversion of aEPEC with a stx-phage in vitro underlines the important role of aEPEC as progenitors of EHEC. Given the high prevalence and the wide host range of aEPEC acceptors, their high risk of zoonotic transmission should be recognized in infection control measures.

Authors+Show Affiliations

Institute for Microbiology and Epizootics, Freie Universität Berlin, Berlin, Germany.Institute for Microbiology and Epizootics, Freie Universität Berlin, Berlin, Germany.Institute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Greifswald - Insel Riems, Germany.Institute of Food Science and Biotechnology, University of Hohenheim, Hohenheim, Germany.Robert Koch Institute, Berlin, Germany.Robert Koch Institute, Wernigerode, Germany.Institute for Microbiology and Epizootics, Freie Universität Berlin, Berlin, Germany.Wellcome Trust Sanger Institute, Hinxton, Cambridge, United Kingdom.Wellcome Trust Sanger Institute, Hinxton, Cambridge, United Kingdom.Institute for Hygiene, University Münster, Münster, Germany.Institute for Microbiology and Epizootics, Freie Universität Berlin, Berlin, Germany; Institute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Greifswald - Insel Riems, Germany. Electronic address: WielerLH@rki.de.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

29937391

Citation

Eichhorn, Inga, et al. "Lysogenic Conversion of Atypical Enteropathogenic Escherichia Coli (aEPEC) From Human, Murine, and Bovine Origin With Bacteriophage Φ3538 Δstx2::cat Proves Their Enterohemorrhagic E. Coli (EHEC) Progeny." International Journal of Medical Microbiology : IJMM, vol. 308, no. 7, 2018, pp. 890-898.
Eichhorn I, Heidemanns K, Ulrich RG, et al. Lysogenic conversion of atypical enteropathogenic Escherichia coli (aEPEC) from human, murine, and bovine origin with bacteriophage Φ3538 Δstx2::cat proves their enterohemorrhagic E. coli (EHEC) progeny. Int J Med Microbiol. 2018;308(7):890-898.
Eichhorn, I., Heidemanns, K., Ulrich, R. G., Schmidt, H., Semmler, T., Fruth, A., Bethe, A., Goulding, D., Pickard, D., Karch, H., & Wieler, L. H. (2018). Lysogenic conversion of atypical enteropathogenic Escherichia coli (aEPEC) from human, murine, and bovine origin with bacteriophage Φ3538 Δstx2::cat proves their enterohemorrhagic E. coli (EHEC) progeny. International Journal of Medical Microbiology : IJMM, 308(7), 890-898. https://doi.org/10.1016/j.ijmm.2018.06.005
Eichhorn I, et al. Lysogenic Conversion of Atypical Enteropathogenic Escherichia Coli (aEPEC) From Human, Murine, and Bovine Origin With Bacteriophage Φ3538 Δstx2::cat Proves Their Enterohemorrhagic E. Coli (EHEC) Progeny. Int J Med Microbiol. 2018;308(7):890-898. PubMed PMID: 29937391.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Lysogenic conversion of atypical enteropathogenic Escherichia coli (aEPEC) from human, murine, and bovine origin with bacteriophage Φ3538 Δstx2::cat proves their enterohemorrhagic E. coli (EHEC) progeny. AU - Eichhorn,Inga, AU - Heidemanns,Katrin, AU - Ulrich,Rainer G, AU - Schmidt,Herbert, AU - Semmler,Torsten, AU - Fruth,Angelika, AU - Bethe,Astrid, AU - Goulding,David, AU - Pickard,Derek, AU - Karch,Helge, AU - Wieler,Lothar H, Y1 - 2018/06/19/ PY - 2018/04/23/received PY - 2018/06/04/revised PY - 2018/06/16/accepted PY - 2018/6/26/pubmed PY - 2019/1/12/medline PY - 2018/6/26/entrez KW - EHEC KW - Lysogenic conversion KW - Microevolution KW - aEPEC KW - stx-converting bacteriophage SP - 890 EP - 898 JF - International journal of medical microbiology : IJMM JO - Int J Med Microbiol VL - 308 IS - 7 N2 - Bacteriophages play an important role in the evolution of bacterial pathogens. A phage-mediated transfer of stx-genes to atypical enteropathogenic E. coli (aEPEC) which are prevalent in different hosts, would convert them to enterohemorrhagic E. coli (EHEC). We decided to confirm this hypothesis experimentally to provide conclusive evidence that aEPEC isolated from different mammalian hosts are indeed progenitors of typical EHEC which gain the ability to produce Shiga-Toxin by lysogeny with stx-converting bacteriophages, utilizing the model phage Φ3538 Δstx2::cat. We applied a modified in vitro plaque-assay, using a high titer of a bacteriophage carrying a deletion in the stx2 gene (Φ3538 Δstx2::cat) to increase the detection of lysogenic conversion events. Three wild-type aEPEC strains were chosen as acceptor strains: the murine aEPEC-strain IMT14505 (sequence type (ST)28, serotype Ont:H6), isolated from a striped field mouse (Apodemus agrarius) in the surrounding of a cattle shed, and the human aEPEC-strain 910#00 (ST28, Ont:H6). The close genomic relationship of both strains implies a high zoonotic potential. A third strain, the bovine aEPEC IMT19981, was of serotype O26:H11 and ST21 (STC29). All three aEPEC were successfully lysogenized with phage Φ3538 Δstx2::cat. Integration of the bacteriophage DNA into the aEPEC host genomes was confirmed by amplification of chloramphenicol transferase (cat) marker gene and by Southern-Blot hybridization. Analysis of the whole genome sequence of each of the three lysogens showed that the bacteriophage was integrated into the known tRNA integration site argW, which is highly variable among E. coli. In conclusion, the successful lysogenic conversion of aEPEC with a stx-phage in vitro underlines the important role of aEPEC as progenitors of EHEC. Given the high prevalence and the wide host range of aEPEC acceptors, their high risk of zoonotic transmission should be recognized in infection control measures. SN - 1618-0607 UR - https://www.unboundmedicine.com/medline/citation/29937391/Lysogenic_conversion_of_atypical_enteropathogenic_Escherichia_coli__aEPEC__from_human_murine_and_bovine_origin_with_bacteriophage_Φ3538_Δstx2::cat_proves_their_enterohemorrhagic_E__coli__EHEC__progeny_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1438-4221(18)30207-8 DB - PRIME DP - Unbound Medicine ER -