Virulence of Shigatoxigenic and Enteropathogenic Escherichia coli O80:H2 in Galleria mellonella Larvae: Comparison of the Roles of the pS88 Plasmids and STX2d Phage
[en] The invasiveness properties of Shigatoxigenic and enteropathogenic Escherichia coli (STEC and EPEC) O80:H2 in humans and calves are encoded by genes located on a pS88-like ColV conjugative plasmid. The main objectives of this study in larvae of the Galleria mellonella moth were therefore to compare the virulence of eight bovine STEC and EPEC O80:H2, of two E. coli pS88 plasmid transconjugant and STX2d phage transductant K12 DH10B, of four E. coli O80:non-H2, and of the laboratory E. coli K12 DH10B strains. Thirty larvae per strain were inoculated in the last proleg with 10 μL of tenfold dilutions of each bacterial culture corresponding to 10 to 106 colony-forming units (CFUs). The larvae were kept at 37 °C and their mortality rate was followed daily for four days. The main results were that: (i) not only the STEC and EPEC O80:H2, but also different E. coli O80:non-H2 were lethal for the larvae at high concentrations (from 104 to 106 CFU) with some variation according to the strain; (ii) the Stx2d toxin and partially the pS88 plasmid were responsible for the lethality caused by the E. coli O80:H2; (iii) the virulence factors of E. coli O80:non-H2 were not identified. The general conclusions are that, although the Galleria mellonella larvae represent a useful first-line model to study the virulence of bacterial pathogens, they are more limited in identifying their actual virulence properties.
Disciplines :
Veterinary medicine & animal health
Author, co-author :
Ikeda, Rie ; Université de Liège - ULiège > Fundamental and Applied Research for Animals and Health (FARAH)
Laforêt, Fanny ; Université de Liège - ULiège > Fundamental and Applied Research for Animals and Health (FARAH) > FARAH: Santé publique vétérinaire
Antoine, Céline ; Université de Liège - ULiège > Fundamental and Applied Research for Animals and Health (FARAH)
Adachi, Mare; Veterinary Bacteriology, Department of Infectious and Parasitic Diseases, Faculty of Veterinary Medicine, Center for Fundamental and Applied Research for Animals and Health (FARAH), University of Liège, B-4000 Liege, Belgium
Nakamura, Keiji ; Department of Bacteriology, Faculty of Medical Science, Kyushu University, Fukuoka 812-8582, Japan
Habets, Audrey ; Université de Liège - ULiège > Faculté de Médecine Vétérinaire > Doct. scienc. vétér.
Kler, Cassandra; Veterinary Bacteriology, Department of Infectious and Parasitic Diseases, Faculty of Veterinary Medicine, Center for Fundamental and Applied Research for Animals and Health (FARAH), University of Liège, B-4000 Liege, Belgium
De Rauw, Klara ; Belgium National Reference Center of STEC (NRC STEC), Universitair Ziekenhuis Brussel (UZ Brussel), Vrije Universiteit Brussel (VUB), B-1090 Brussels, Belgium
Hayashi, Tetsuya; Department of Bacteriology, Faculty of Medical Science, Kyushu University, Fukuoka 812-8582, Japan
Mainil, Jacques ✱; Université de Liège - ULiège > Département des maladies infectieuses et parasitaires (DMI)
Thiry, Damien ✱; Université de Liège - ULiège > Département des maladies infectieuses et parasitaires (DMI) > Bactériologie vétérinaire et maladies bactériennes animales
✱ These authors have contributed equally to this work.
Language :
English
Title :
Virulence of Shigatoxigenic and Enteropathogenic Escherichia coli O80:H2 in Galleria mellonella Larvae: Comparison of the Roles of the pS88 Plasmids and STX2d Phage
Russo T.A. Johnson J.R. Proposal for a New Inclusive Designation for Extraintestinal Pathogenic Isolates of Escherichia coli: ExPEC J. Infect. Dis. 2000 181 1753 1754 10.1086/315418 10823778
Tozzoli R. Scheutz F. Diarrheagenic Escherichia coli Infections in Humans Pathogenic Escherichia coli: Molecular and Cellular Microbiology Morabito S. Caister Academic Press Norfolk, UK 2014 1 18 978-1-908230-37-9
Mainil J.G. Fairbrother M.J. Pathogenic Escherichia coli in Domestic Mammals and Birds Pahogenic Escherichia coli: Molecular and Cellular Microbiology Morabito S. Caister Academic Press Norfolk, UK 2014 19 43 978-1-908230-37-9
Borgatta B. Kmet-Lunaček N. Rello J.E. coli O104:H4 Outbreak and Haemolytic-Uraemic Syndrome Med. Intensiv. 2012 36 576 583 10.1016/j.medin.2011.11.022 22244215
Piérard D. De Greve H. Haesebrouck F. Mainil J. O157:H7 and O104:H4 Vero/Shiga Toxin-Producing Escherichia coli Outbreaks: Respective Role of Cattle and Humans Vet. Res. 2012 43 13 10.1186/1297-9716-43-13
EFSA BIOHAZ Panel Koutsoumanis K. Allende A. Alvarez-Ordóñez A. Bover-Cid S. Chemaly M. Davies R. De Cesare A. Herman L. Hilbert F. et al. Pathogenicity Assessment of Shiga Toxin-Producing Escherichia coli (STEC) and the Public Health Risk Posed by Contamination of Food with STEC EFSA J. 2020 18 5967 10.2903/j.efsa.2019.5898
Bai X. Fu S. Zhang J. Fan R. Xu Y. Sun H. He X. Xu J. Xiong Y. Identification and Pathogenomic Analysis of an Escherichia coli Strain Producing a Novel Shiga Toxin 2 Subtype Sci. Rep. 2018 8 6756 10.1038/s41598-018-25233-x
Gill A. Dussault F. McMahon T. Petronella N. Wang X. Cebelinski E. Scheutz F. Weedmark K. Blais B. Carrillo C. Characterization of Atypical Shiga Toxin Gene Sequences and Description of Stx2j, a New Subtype J. Clin. Microbiol. 2022 60 e0222921 10.1128/jcm.02229-21
Martin C.C. Svanevik C.S. Lunestad B.T. Sekse C. Johannessen G.S. Isolation and Characterisation of Shiga Toxin-Producing Escherichia coli from Norwegian Bivalves Food Microbiol. 2019 84 103268 10.1016/j.fm.2019.103268
Yang X. Bai X. Zhang J. Sun H. Fu S. Fan R. He X. Scheutz F. Matussek A. Xiong Y. Escherichia coli Strains Producing a Novel Shiga Toxin 2 Subtype Circulate in China Int. J. Med. Microbiol. 2020 310 151377 10.1016/j.ijmm.2019.151377
Karmali M.A. Mascarenhas M. Shen S. Ziebell K. Johnson S. Reid-Smith R. Isaac-Renton J. Clark C. Rahn K. Kaper J.B. Association of Genomic O Island 122 of Escherichia coli EDL 933 with Verocytotoxin-Producing Escherichia coli Seropathotypes That Are Linked to Epidemic and/or Serious Disease J. Clin. Microbiol. 2003 41 4930 4940 10.1128/JCM.41.11.4930-4940.2003
Soysal N. Mariani-Kurkdjian P. Smail Y. Liguori S. Gouali M. Loukiadis E. Fach P. Bruyand M. Blanco J. Bidet P. et al. Enterohemorrhagic Escherichia coli Hybrid Pathotype O80:H2 as a New Therapeutic Challenge Emerg. Infect. Dis. 2016 22 1604 1612 10.3201/eid2209.160304 27533474
Nüesch-Inderbinen M. Morach M. Cernela N. Althaus D. Jost M. Mäusezahl M. Bloomberg G. Stephan R. Serotypes and Virulence Profiles of Shiga Toxin-Producing Escherichia coli Strains Isolated during 2017 from Human Infections in Switzerland Int. J. Med. Microbiol. 2018 308 933 939 10.1016/j.ijmm.2018.06.011 30042042
Cointe A. Bizot E. Delannoy S. Fach P. Bidet P. Birgy A. Weill F.-X. Lefèvre S. Mariani-Kurkdjian P. Bonacorsi S. Emergence of New ST301 Shiga Toxin-Producing Escherichia coli Clones Harboring Extra-Intestinal Virulence Traits in Europe Toxins 2021 13 686 10.3390/toxins13100686 34678979
Gigliucci F. van Hoek A.H.A.M. Chiani P. Knijn A. Minelli F. Scavia G. Franz E. Morabito S. Michelacci V. Genomic Characterization of HlyF-Positive Shiga Toxin-Producing Escherichia coli, Italy and the Netherlands, 2000–2019 Emerg. Infect. Dis. 2021 27 853 861 10.3201/eid2703.203110 33622476
Wijnsma K.L. Schijvens A.M. Rossen J.W.A. Kooistra-Smid A.M.D. Schreuder M.F. van de Kar N.C.A.J. Unusual Severe Case of Hemolytic Uremic Syndrome Due to Shiga Toxin 2d-Producing E. coli O80:H2 Pediatr. Nephrol. 2017 32 1263 1268 10.1007/s00467-017-3642-3
De Rauw K. Thiry D. Caljon B. Saulmont M. Mainil J. Piérard D. Characteristics of Shiga toxin Producing- and Enteropathogenic Escherichia coli of the Emerging Serotype O80:H2 Isolated from Humans and Diarrheic Calves in Belgium Clin. Microbiol. Infect. 2019 25 111.e5 111.e8 10.1016/j.cmi.2018.07.023
Peigne C. Bidet P. Mahjoub-Messai F. Plainvert C. Barbe V. Médigue C. Frapy E. Nassif X. Denamur E. Bingen E. et al. The Plasmid of Escherichia coli Strain S88 (O45:K1:H7) That Causes Neonatal Meningitis Is Closely Related to Avian Pathogenic E. coli Plasmids and Is Associated with High-Level Bacteremia in a Neonatal Rat Meningitis Model Infect. Immun. 2009 77 2272 2284 10.1128/IAI.01333-08
Thiry D. Saulmont M. Takaki S. De Rauw K. Duprez J.-N. Iguchi A. Piérard D. Mainil J.G. Enteropathogenic Escherichia coli O80:H2 in Young Calves with Diarrhea, Belgium Emerg. Infect. Dis. 2017 23 2093 2095 10.3201/eid2312.170450
Habets A. Crombé F. Nakamura K. Guérin V. De Rauw K. Piérard D. Saulmont M. Hayashi T. Mainil J.G. Thiry D. Genetic Characterization of Shigatoxigenic and Enteropathogenic Escherichia coli O80:H2 from Diarrhoeic and Septicaemic Calves and Relatedness to Human Shigatoxigenic E. coli O80:H2 J. Appl. Microbiol. 2021 130 258 264 10.1111/jam.14759
Cointe A. Birgy A. Mariani-Kurkdjian P. Liguori S. Courroux C. Blanco J. Delannoy S. Fach P. Loukiadis E. Bidet P. et al. Emerging Multidrug-Resistant Hybrid Pathotype Shiga Toxin-Producing Escherichia coli O80 and Related Strains of Clonal Complex 165, Europe Emerg. Infect. Dis. 2018 24 2262 2269 10.3201/eid2412.180272
Lewis D.I. Animal Experimentation: Implementation and Application of the 3Rs Emerg. Top Life Sci. 2019 3 675 679 10.1042/ETLS20190061
Harnish J.M. Link N. Yamamoto S. Drosophila as a Model for Infectious Diseases Int. J. Mol. Sci. 2021 22 2724 10.3390/ijms22052724 33800390
Ahlawat S. Sharma K.K. Lepidopteran Insects: Emerging Model Organisms to Study Infection by Enteropathogens Folia Microbiol. 2023 68 181 196 10.1007/s12223-022-01014-y 36417090
Cutuli M.A. Petronio Petronio G. Vergalito F. Magnifico I. Pietrangelo L. Venditti N. Di Marco R. Galleria mellonella as a Consolidated in vivo Model Hosts: New Developments in Antibacterial Strategies and Novel Drug Testing Virulence 2019 10 527 541 10.1080/21505594.2019.1621649 31142220
Pereira M.F. Rossi C.C. da Silva G.C. Rosa J.N. Bazzolli D.M.S. Galleria mellonella as an Infection Model: An In-Depth Look at why it Works and Practical Considerations for Successful Application Pathog. Dis. 2020 78 ftaa056 10.1093/femspd/ftaa056 32960263
Antoine C. Laforêt F. Blasdel B. Fall A. Duprez J.-N. Mainil J. Delcenserie V. Thiry D. In vitro Characterization and in vivo Efficacy Assessment in Galleria mellonella Larvae of Newly Isolated Bacteriophages against Escherichia coli K1 Viruses 2021 13 2005 10.3390/v13102005 34696434
Wojda I. Staniec B. Sułek M. Kordaczuk J. The Greater Wax Moth Galleria mellonella: Biology and Use in Immune Studies Pathog. Dis. 2020 78 ftaa057 10.1093/femspd/ftaa057
Habets A. Antoine C. Wagemans J. Vermeersch M. Laforêt F. Diderich J. Lavigne R. Mainil J. Thiry D. Impact of Shiga-Toxin Encoding Gene Transduction from O80:H2 Shiga Toxigenic Escherichia coli (STEC) on Non-STEC Strains Sci. Rep. 2022 12 21587 10.1038/s41598-022-26198-8
Chen R.Y. Keddie B.A. The Galleria mellonella-Enteropathogenic Escherichia coli Model System: Characterization of Pathogen Virulence and Insect Immune Responses J. Insect. Sci. 2021 21 7 10.1093/jisesa/ieab046
Morgan J.K. Ortiz J.A. Riordan J.T. The Role for TolA in Enterohemorrhagic Escherichia coli Pathogenesis and Virulence Gene Transcription Microb. Pathog. 2014 77 42 52 10.1016/j.micpath.2014.10.010
De la Cruz M.A. Morgan J.K. Ares M.A. Yanez-Santos J.A. Riordan J.T. Giron J.A. The Two-Component Sytem CpxRA Negatively Regulates the Locus of Enterocyte Effacement of Enterohemorrhagic Escherichia coli Involving σ32 and Lon Protease Front. Cell. Infect. Microbiol. 2016 6 11 10.3389/fcimb.2016.00011 26904510
Ikeda R. Nakamura K. Saulmont M. Habets A. Duprez J.-N. Korsak N. Hayashi T. Thiry D. Mainil J.G. Escherichia coli O80 in Healthy Cattle: Absence of Shigatoxigenic and Enteropathogenic E. coli O80:H2 and (Phylo) Genomics of Non-Clonal Complex 165 E. coli O80 Microorganisms 2023 11 230 10.3390/microorganisms11020230 36838195
Liu D. Reeves P.R. Escherichia coli K12 Regains Its O Antigen Microbiology 1994 140 Pt 1 49 57 10.1099/13500872-140-1-49 7512872
Gilson L. Mahanty H.K. Kolter R. Four Plasmid Genes Are Required for Colicin V Synthesis, Export, and Immunity J. Bacteriol. 1987 169 2466 2470 10.1128/jb.169.6.2466-2470.1987
Bauer A.W. Kirby W.M. Sherris J.C. Turck M. Antibiotic Susceptibility Testing by a Standardized Single Disk Method Am. J. Clin. Pathol. 1966 45 493 496 10.1093/ajcp/45.4_ts.493
Pohl P. Introduction à la Plasmidologie: Données actuelles sur sa Signification en Thérapeutique et en Pathologie J. Pharm. Belg. 1981 36 165 174
Brosius J. Holy A. Regulation of Ribosomal RNA Promoters with a Synthetic Lac Operator Proc. Natl. Acad. Sci. USA 1984 81 6929 6933 10.1073/pnas.81.22.6929
Iguchi A. Iyoda S. Seto K. Morita-Ishihara T. Scheutz F. Ohnishi M. Pathogenic E. coli Working Group in Japan Escherichia coli O-Genotyping PCR: A Comprehensive and Practical Platform for Molecular O Serogrouping J. Clin. Microbiol. 2015 53 2427 2432 10.1128/JCM.00321-15
Banjo M. Iguchi A. Seto K. Kikuchi T. Harada T. Scheutz F. Iyoda S. Pathogenic E. coli Working Group in Japan Escherichia coli H-Genotyping PCR: A Complete and Practical Platform for Molecular H Typing J. Clin. Microbiol. 2018 56 e00190-18 10.1128/JCM.00190-18
Vandekerchove D. Vandemaele F. Adriaensen C. Zaleska M. Hernalsteens J.-P. De Baets L. Butaye P. Van Immerseel F. Wattiau P. Laevens H. et al. Virulence-Associated Traits in Avian Escherichia coli: Comparison between Isolates from Colibacillosis-Affected and Clinically Healthy Layer Flocks Vet. Microbiol. 2005 108 75 87 10.1016/j.vetmic.2005.02.009
Johnson T.J. Siek K.E. Johnson S.J. Nolan L.K. DNA Sequence of a ColV Plasmid and Prevalence of Selected Plasmid-Encoded Virulence genes among Avian Escherichia coli Strains J. Bacteriol. 2006 188 755 758 10.1128/JB.188.2.745-758.2006 16385064
Ling J. Pan H. Gao Q. Xiong L. Zhou Y. Zhang D. Gao S. Liu X. Aerobactin Synthesis Genes iucA and iucC Contribute to the Pathogenicity of Avian Pathogenic Escherichia coli O2 Strain E058 PLoS ONE 2013 8 e57794 10.1371/journal.pone.0057794 23460907
Braun V. Helbig S. Patzer S.I. Pramanik A. Römer C. Import and Export of Bacterial Protein Toxins Int. J. Med. Microbiol. 2015 305 238 242 10.1016/j.ijmm.2014.12.006 25620353
Tarr P.I. Bilge S.S. Vary J.C. Jr. Jelacic S. Habeeb R.L. Ward T.R. Baylor M.R. Besser T.E. Iha: A Novel Escherichia coli O157:H7 Adherence-conferring Molecule Encoded on a Recently Acquired Chromosomal Island of Conserved Structure Infect. Immun. 2000 68 1400 1407 10.1128/IAI.68.3.1400-1407.2000
Schmidt H. Hensel M. Pathogenicity Islands in Bacterial Pathogenesis Clin. Microbiol. Rev. 2004 17 14 56 10.1128/CMR.17.1.14-56.2004
Mellata M. Dho-Moulin M. Dozois C.M. Curtiss R. Brown P.K. Arné P. Brée A. Desautels C. Fairbrother J.M. Role of Virulence Factors in Resistance of Avian Pathogenic Escherichia coli to Serum and in Pathogenicity Infect. Immun. 2003 71 536 540 10.1128/IAI.71.1.536-540.2003
Caza M. Lépine F. Milot S. Dozois C.M. Specific Roles of the iroBCDEN Genes in Virulence of an Avian Pathogenic Escherichia coli O78 Strain and in Production of Salmochelins Infect. Immun. 2008 76 3539 3549 10.1128/IAI.00455-08
Gyles C. Boerlin P. Horizontally Transferred Genetic Elements and their Role in Pathogenesis of Bacterial Disease Vet. Pathol. 2014 51 328 340 10.1177/0300985813511131
Datsenko K.A. Wanner B.L. One-Step Inactivation of Chromosomal Genes in Escherichia coli K-12 Using PCR Products Proc. Natl. Acad. Sci. USA 2000 97 6640 6645 10.1073/pnas.120163297
Hutchens M. Luker G.D. Applications of Bioluminescence Imaging to the Study of Infectious Diseases Cell. Microbiol. 2007 9 2315 2322 10.1111/j.1462-5822.2007.00995.x
Witcomb L.A. Czupryna J. Francis K.P. Frankel G. Taylor P.W. Non-Invasive Three-Dimensional Imaging of Escherichia coli K1 Infection Using Diffuse Light Imaging Tomography Combined with Micro-Computed Tomography Methods 2017 127 62 68 10.1016/j.ymeth.2017.05.005 28522324