Enterohemorrhagic Escherichia coli in human infection: in vivo evolution of a bacterial pathogen.

BACKGROUND Enterohemorrhagic Escherichia coli (EHEC) cause most cases of the hemolytic uremic syndrome (HUS) worldwide. To investigate genetic changes in EHEC during the course of human infection, we analyzed consecutive stool samples and shed isolates from patients with HUS, focusing on the genes encoding Shiga toxin (stx) and intimin (eae). METHODS Sequential stool samples from 210 patients with HUS were investigated for the persistence of E. coli strains harboring stx and/or eae. Initial stool samples were collected during the acute phase of HUS, and subsequent stool samples were collected 3-16 days later (median interval, 8 days). RESULTS Organisms that were stx and eae positive (stx+/eae+ strains; n=137) or stx negative and eae positive (stx-/eae+ strains; n=5) were detected in the initial stool samples from 142 patients. Subsequently, the proportion of those who shed stx+/eae+ strains decreased to 13 of 210 patients, whereas the proportion of those who shed strains that were stx-/eae+ increased to 12 of 210 patients. Seven patients who initially excreted strains that were stx+/eae+ shed, at second analysis, stx-/eae+ strains of the same serotypes; they had no free fecal Shiga toxin at follow-up. Comparison of the initial and follow-up isolates from these patients with use of molecular-epidemiological methods revealed loss of stx genes and genomic rearrangement. CONCLUSIONS We demonstrate the loss of a critical bacterial virulence factor from pathogens during very brief intervals in the human host. These genetic changes have evolutionary, diagnostic, and clinical implications. Generation of stx- mutants might contribute to subclonal evolution and evolutionary success.

[1]  P. Tarr,et al.  Shiga-toxin-producing and haemolytic uraemic syndrome , 2005 .

[2]  M. Schmidt,et al.  Comparative Analysis of Locus of Enterocyte Effacement Pathogenicity Islands of Atypical Enteropathogenic Escherichia coli , 2004, Infection and Immunity.

[3]  A. Mellmann,et al.  Phylogeny, Clinical Associations, and Diagnostic Utility of the Pilin Subunit Gene (sfpA) of Sorbitol-Fermenting, Enterohemorrhagic Escherichia coli O157:H− , 2004, Journal of Clinical Microbiology.

[4]  M. Sinclair,et al.  Escherichia coli and Community-acquired Gastroenteritis, Melbourne, Australia , 2004, Emerging infectious diseases.

[5]  Herbert Schmidt,et al.  Shiga toxin-encoding bacteriophages--genomes in motion. , 2004, International journal of medical microbiology : IJMM.

[6]  Ulrich Dobrindt,et al.  Genomic islands in pathogenic and environmental microorganisms , 2004, Nature Reviews Microbiology.

[7]  H. Karch,et al.  Phenotypic and Genotypic Analyses of Enterohemorrhagic Escherichia coli O145 Strains from Patients in Germany , 2004, Journal of Clinical Microbiology.

[8]  L. Beutin,et al.  HEp-2 Cell Adherence, Actin Aggregation, and Intimin Types of Attaching and Effacing Escherichia coli Strains Isolated from Healthy Infants in Germany and Australia , 2003, Infection and Immunity.

[9]  Phillip I. Tarr,et al.  Escherichia coli O157:H7 Shiga Toxin-Encoding Bacteriophages: Integrations, Excisions, Truncations, and Evolutionary Implications , 2003, Journal of bacteriology.

[10]  H. Karch,et al.  Cytolethal Distending Toxin Gene Cluster in Enterohemorrhagic Escherichiacoli O157:H− and O157:H7: Characterization and Evolutionary Considerations , 2003, Infection and Immunity.

[11]  A. Friedrich,et al.  Shiga Toxin 1c-Producing Escherichia coli Strains: Phenotypic and Genetic Characterization and Association with Human Disease , 2003, Journal of Clinical Microbiology.

[12]  H. Karch,et al.  Cluster of hemolytic-uremic syndrome caused by Shiga toxin-producing Escherichia coli O26:H11 , 2003, The Pediatric infectious disease journal.

[13]  A. Edefonti,et al.  Shiga Toxin–Producing Escherichia coli Infections Associated with Hemolytic Uremic Syndrome, Italy, 1988–2000 , 2003, Emerging infectious diseases.

[14]  A. Fruth,et al.  Subtyping of pathogenic Escherichia coli strains using flagellar (H)-antigens: serotyping versus fliC polymorphisms. , 2003, International journal of medical microbiology : IJMM.

[15]  L. Beutin,et al.  Genetic Diversity of Intimin Genes of Attaching and Effacing Escherichia coli Strains , 2002, Journal of Clinical Microbiology.

[16]  H. Karch,et al.  Clinical course and the role of shiga toxin-producing Escherichia coli infection in the hemolytic-uremic syndrome in pediatric patients, 1997-2000, in Germany and Austria: a prospective study. , 2002, The Journal of infectious diseases.

[17]  U. Buchholz,et al.  A multistate outbreak of Shiga toxin-producing Escherichia coli O26:H11 infections in Germany, detected by molecular subtyping surveillance. , 2002, The Journal of infectious diseases.

[18]  W. Rabsch,et al.  Clonal diversity of Shiga toxin-producing Escherichia coli O103:H2/H(-) in Germany. , 2002, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[19]  H. Karch,et al.  Identification, Characterization, and Distribution of a Shiga Toxin 1 Gene Variant (stx1c) in Escherichia coli Strains Isolated from Humans , 2002, Journal of Clinical Microbiology.

[20]  A. Friedrich,et al.  Escherichia coli harboring Shiga toxin 2 gene variants: frequency and association with clinical symptoms. , 2002, The Journal of infectious diseases.

[21]  D. Falush,et al.  Recombination and mutation during long-term gastric colonization by Helicobacter pylori: Estimates of clock rates, recombination size, and minimal age , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[22]  A. Siitonen,et al.  Clinical Isolates of Non-O157 Shiga Toxin-ProducingEscherichia coli: Serotypes, Virulence Characteristics, and Molecular Profiles of Strains of the Same Serotype , 2001, Journal of Clinical Microbiology.

[23]  E. Elliott,et al.  Nationwide study of haemolytic uraemic syndrome: clinical, microbiological, and epidemiological features , 2001, Archives of disease in childhood.

[24]  A. Abe,et al.  Isogenic Strain of Escherichia coliO157:H7 That Has Lost both Shiga Toxin 1 and 2 Genes , 2001, Clinical Diagnostic Laboratory Immunology.

[25]  H. Karch,et al.  Novel Type of Fimbriae Encoded by the Large Plasmid of Sorbitol-Fermenting Enterohemorrhagic Escherichia coli O157:H− , 2001, Infection and Immunity.

[26]  H. Karch.  The Role of Virulence Factors in Enterohemorrhagic Escherichia coli (EHEC) - Associated Hemolytic-Uremic Syndrome , 2001, Seminars in thrombosis and hemostasis.

[27]  J. H. Green,et al.  The United States National Prospective Hemolytic Uremic Syndrome Study: microbiologic, serologic, clinical, and epidemiologic findings. , 2001, The Journal of infectious diseases.

[28]  U. Alon.  The Risk of the Hemolytic-Uremic Syndrome After Antibiotic Treatment of Escherichia coli 0157:H7 Infections , 2001 .

[29]  M. Waldor,et al.  Human Neutrophils and Their Products Induce Shiga Toxin Production by Enterohemorrhagic Escherichia coli , 2001, Infection and Immunity.

[30]  Craig S. Wong,et al.  The risk of the hemolytic-uremic syndrome after antibiotic treatment of Escherichia coli O157:H7 infections. , 2000, The New England journal of medicine.

[31]  A. Caprioli,et al.  Typing of Intimin Genes in Human and Animal Enterohemorrhagic and Enteropathogenic Escherichia coli: Characterization of a New Intimin Variant , 2000, Infection and Immunity.

[32]  H. Karch,et al.  Molecular characteristics and epidemiological significance of Shiga toxin-producing Escherichia coli O26 strains. , 2000, Journal of clinical microbiology.

[33]  T. Honda,et al.  Induction of Prophages of EnterohemorrhagicEscherichia coli O157:H7 with Norfloxacin , 1999, Journal of bacteriology.

[34]  M. Petric,et al.  Evaluation of a Microplate Latex Agglutination Method (Verotox-F Assay) for Detecting and Characterizing Verotoxins (Shiga Toxins) in Escherichia coli , 1999, Journal of Clinical Microbiology.

[35]  H. Karch,et al.  Non-O157:H7 pathogenic Shiga toxin-producing Escherichia coli: phenotypic and genetic profiling of virulence traits and evidence for clonality. , 1999, The Journal of infectious diseases.

[36]  James P. Nataro,et al.  Diarrheagenic Escherichia coli , 1998, Clinical Microbiology Reviews.

[37]  D H Persing,et al.  Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing , 1995, Journal of clinical microbiology.

[38]  L. Beutin,et al.  Molecular analysis of the plasmid-encoded hemolysin of Escherichia coli O157:H7 strain EDL 933 , 1995, Infection and immunity.

[39]  W. L. Payne,et al.  Simultaneous identification of strains of Escherichia coli serotype O157:H7 and their Shiga-like toxin type by mismatch amplification mutation assay-multiplex PCR , 1995, Journal of clinical microbiology.

[40]  H. Karch,et al.  Prevalence of attaching and effacing Escherichia coli in stool samples from patients and controls. , 1994, Zentralblatt fur Bakteriologie : international journal of medical microbiology.

[41]  M. McKee,et al.  The role of the eae gene of enterohemorrhagic Escherichia coli in intimate attachment in vitro and in a porcine model. , 1993, The Journal of clinical investigation.

[42]  J. Heesemann,et al.  Frequent loss of Shiga-like toxin genes in clinical isolates of Escherichia coli upon subcultivation , 1992, Infection and immunity.

[43]  J. Yu,et al.  Cloning and characterization of the eae gene of enterohaemorrhagic Escherichia coli O157:H7 , 1992, Molecular microbiology.

[44]  H. Karch,et al.  High incidence of serum antibodies to Escherichia coli O157 lipopolysaccharide in children with hemolytic-uremic syndrome. , 1991, The Journal of pediatrics.

[45]  J. Yu,et al.  A genetic locus of enteropathogenic Escherichia coli necessary for the production of attaching and effacing lesions on tissue culture cells. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[46]  P. Tarr,et al.  Escherichia coli 0157:H7 and the Hemolytic Uremic Syndrome: Importance of Early Cultures in Establishing the Etiology , 1990 .

[47]  L. Beutin,et al.  Close association of verotoxin (Shiga-like toxin) production with enterohemolysin production in strains of Escherichia coli , 1989, Journal of clinical microbiology.

[48]  L. Becker,et al.  The histopathology of the hemolytic uremic syndrome associated with verocytotoxin-producing Escherichia coli infections. , 1988, Human pathology.

[49]  R. Holmes,et al.  Shiga-like toxin-converting phages from Escherichia coli strains that cause hemorrhagic colitis or infantile diarrhea. , 1984, Science.

[50]  S. Formal,et al.  ESCHERICHIA COLI 0157:H7 STRAINS ASSOCIATED WITH HAEMORRHAGIC COLITIS IN THE UNITED STATES PRODUCE A SHIGELLA DYSENTERIAE 1 (SHIGA) LIKE CYTOTOXIN , 1983, The Lancet.