No evidence of the Shiga toxin-producing E. coli O104:H4 outbreak strain or enteroaggregative E. coli (EAEC) found in cattle faeces in northern Germany, the hotspot of the 2011 HUS outbreak area

Gut Pathogens

Astrid Bethe²  Sebastian Guenther²  Helge Karch¹  Lutz Geue³  Bianca Kinnemann²  Esther M Antao²  Inga Eichhorn²  Torsten Semmler²  Lothar H Wieler² 

[1] Institute for Hygiene and the National Consulting Laboratory for Haemolytic Uraemic Syndrome, University of Münster, Germany;Institute of Microbiology and Epizootics, Veterinary Faculty, Freie Universität Berlin, Germany;Institute of Epidemiology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Seestrasse 55, D-16868 Wusterhausen, Germany

关键词: outbreak;    cattle;    HUS;    O104:H4;    enteroaggregative;    EAEC;    E.coli;    Shiga toxin;   

Others  :  833646
DOI  :  10.1186/1757-4749-3-17

 received in 2011-10-13, accepted in 2011-11-03,  发布年份 2011

【 摘 要 】

Background

Ruminants, in particular bovines, are the primary reservoir of Shiga toxin-producing E. coli (STEC), but whole genome analyses of the current German ESBL-producing O104:H4 outbreak strain of sequence type (ST) 678 showed this strain to be highly similar to enteroaggregative E. coli (EAEC). Strains of the EAEC pathotype are basically adapted to the human host. To clarify whether in contrast to this paradigm, the O104:H4 outbreak strain and/or EAEC may also be able to colonize ruminants, we screened a total of 2.000 colonies from faecal samples of 100 cattle from 34 different farms - all located in the HUS outbreak region of Northern Germany - for genes associated with the O104:H4 HUS outbreak strain (stx2, terD, rfb_O104, fliC_H4), STEC (stx1, stx2, escV), EAEC (pAA, aggR, astA), and ESBL-production (bla_CTX-M, bla_TEM, bla_SHV).

Results

The faecal samples contained neither the HUS outbreak strain nor any EAEC. As the current outbreak strain belongs to ST678 and displays an en-teroaggregative and ESBL-producing phenotype, we additionally screened selected strains for ST678 as well as the aggregative adhesion pattern in HEp-2 cells. However, we were unable to find any strains belonging to ST678 or showing an aggregative adhesion pattern. A high percentage of animals (28%) shed STEC, corroborating previous knowl-edge and thereby proving the validity of our study. One of the STEC also harboured the LEE pathogenicity island. In addition, eleven animals shed ESBL-producing E. coli.

Conclusions

While we are aware of the limitations of our survey, our data support the theory, that, in contrast to other Shiga-toxin producing E. coli, cattle are not the reservoir for the O104:H4 outbreak strain or other EAEC, but that the outbreak strain seems to be adapted to humans or might have yet another reservoir, raising new questions about the epidemiology of STEC O104:H4.

【 授权许可】

   
2011 Wieler et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

【 参考文献 】

• [1]Mellmann A, Bielaszewska M, Kock R, Friedrich AW, Fruth A, Middendorf B, Harmsen D, Schmidt MA, Karch H: Analysis of collection of hemolytic uremic syndrome-associated enterohemorrhagic Escherichia coli. Emerg Infect Dis 2008, 14:1287-1290.
• [2]Askar M, Faber M, Frank C, Bernard H, Gilsdorf A, Fruth A, Prager R, Hohle M, Suess T, Wadl M, et al.: Update on the ongoing outbreak of haemolytic uraemic syndrome due to Shiga toxin-producing Escherichia coli (STEC) serotype O104, Germany, May 2011. Euro Surveill 2011., 16
• [3]Bielaszewska M, Mellmann A, Zhang W, Köck R, Fruth A, Bauwens A, Peters G, Karch H: Characterization of the E. coli strain associated with an outbreak of haemolytic uraemic syndrome in Germany, 2011: a microbiological study. Lancet Infect Dis 2011.
• [4]Frank C, Werber D, Cramer JP, Askar M, Faber M, Heiden MA, Bernard H, Fruth A, Prager R, Spode A, et al.: Epidemic Profile of Shiga-Toxin-Producing Escherichia coli O104:H4 Outbreak in Germany - Preliminary Report. N Engl J Med 2011.
• [5]Brzuszkiewicz E, Thurmer A, Schuldes J, Leimbach A, Liesegang H, Meyer FD, Boelter J, Petersen H, Gottschalk G, Daniel R: Genome sequence analyses of two isolates from the recent Escherichia coli outbreak in Germany reveal the emergence of a new pathotype: Entero-Aggregative-Haemorrhagic Escherichia coli (EAHEC). Archives of microbiology 2011.
• [6]Mossoro C, Glaziou P, Yassibanda S, Lan NT, Bekondi C, Minssart P, Bernier C, Le Bouguenec C, Germani Y: Chronic diarrhea, hemorrhagic colitis, and hemolytic-uremic syndrome associated with HEp-2 adherent Escherichia coli in adults infected with human immunodeficiency virus in Bangui, Central African Republic. J Clin Microbiol 2002, 40:3086-3088.
• [7]Mellmann AHD, Cummings CA, Zentz MB, Leopold SR, Rico A, Prior K, Szczepanowski R, Ji Y, Zhang W, McLaughlin SF, Henkhaus JK, Leopold B, Bielaszewska M, Prager R, Brzoska PM, Moore RL, Guenther S, Rothberg JM, Karch H: Prospective genomic characterization of the German enterohemorrhagic Escherichia coli O104:H4 outbreak by rapid next generation sequencing technology. PLoS One 2011.
• [8]Rasko DA, Webster DR, Sahl JW, Bashir A, Boisen N, Scheutz F, Paxinos EE, Sebra R, Chin CS, Iliopoulos D, et al.: Origins of the E. coli Strain Causing an Outbreak of Hemolytic-Uremic Syndrome in Germany. N Engl J Med 2011.
• [9]Cassar CA, Ottaway M, Paiba GA, Futter R, Newbould S, Woodward MJ: Absence of enteroaggregative Escherichia coli in farmed animals in Great Britain. Vet Rec 2004, 154:237-239.
• [10]Uber AP, Trabulsi LR, Irino K, Beutin L, Ghilardi AC, Gomes TA, Liberatore AM, de Castro AF, Elias WP: Enteroaggregative Escherichia coli from humans and animals differ in major phenotypical traits and virulence genes. FEMS Microbiol Lett 2006, 256:251-257.
• [11]Veilleux S, Dubreuil JD: Presence of Escherichia coli carrying the EAST1 toxin gene in farm animals. Vet Res 2006, 37:3-13.
• [12]Okeke IN, Nataro JP: Enteroaggregative Escherichia coli. Lancet Infect Dis 2001, 1:304-313.
• [13]Okeke IN, Wallace-Gadsden F, Simons HR, Matthews N, Labar AS, Hwang J, Wain J: Multi-locus sequence typing of enteroaggregative Escherichia coli isolates from Nigerian children uncovers multiple lineages. PLoS One 2010, 5:e14093.
• [14]Frank C, Faber M, Askar M, Bernard H, Fruth A, Gilsdorf A, Hohle M, Karch H, Krause G, Prager R, et al.: Large and ongoing outbreak of haemolytic uraemic syndrome, Germany, May 2011. Euro Surveill 2011., 16
• [15]Harrington SM, Dudley EG, Nataro JP: Pathogenesis of enteroaggregative Escherichia coli infection. FEMS Microbiol Lett 2006, 254:12-18.
• [16]Beutin L, Geier D, Zimmermann S, Karch H: Virulence markers of Shiga-like toxin-producing Escherichia coli strains originating from healthy domestic animals of different species. J Clin Microbiol 1995, 33:631-635.
• [17]Menrath A, Wieler LH, Heidemanns K, Semmler T, Fruth A, Kemper N: Shiga toxin producing Escherichia coli: identification of non-O157:H7-Super-Shedding cows and related risk factors. Gut Pathog 2010, 2:7.
• [18]Wieler LH, Sobjinski G, Schlapp T, Failing K, Weiss R, Menge C, Baljer G: Longitudinal prevalence study of diarrheagenic Escherichia coli in dairy calves. Berl Munch Tierarztl Wochenschr 2007, 120:296-306.
• [19]Dopfer D, Geue L, de Bree J, de Jong MC: Dynamics of verotoxin-producing Escherichia coli isolated from German beef cattle between birth and slaughter. Prev Vet Med 2006, 73:229-240.
• [20]Geue L, Segura-Alvarez M, Conraths FJ, Kuczius T, Bockemuhl J, Karch H, Gallien P: A long-term study on the prevalence of shiga toxin-producing Escherichia coli (STEC) on four German cattle farms. Epidemiol Infect 2002, 129:173-185.
• [21]Carattoli A: Animal reservoirs for extended spectrum beta-lactamase producers. Clin Microbiol Infect 2008, 14(Suppl 1):117-123.
• [22]Dolejska M, Jurcickova Z, Literak I, Pokludova L, Bures J, Hera A, Kohoutova L, Smola J, Cizek A: IncN plasmids carrying bla CTX-M-1 in Escherichia coli isolates on a dairy farm. Vet Microbiol 2011, 149:513-516.
• [23]Geser N, Stephan R, Kuhnert P, Zbinden R, Kaeppeli U, Cernela N, Haechler H: Fecal carriage of extended-spectrum beta-lactamase-producing Enterobacteriaceae in swine and cattle at slaughter in Switzerland. J Food Prot 2011, 74:446-449.
• [24]Jouini A, Vinue L, Slama KB, Saenz Y, Klibi N, Hammami S, Boudabous A, Torres C: Characterization of CTX-M and SHV extended-spectrum beta-lactamases and associated resistance genes in Escherichia coli strains of food samples in Tunisia. J Antimicrob Chemother 2007, 60:1137-1141.
• [25]Vial PA, Robins-Browne R, Lior H, Prado V, Kaper JB, Nataro JP, Maneval D, Elsayed A, Levine MM: Characterization of enteroadherent-aggregative Escherichia coli, a putative agent of diarrheal disease. J Infect Dis 1988, 158:70-79.
• [26]CLSI: Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals; approved standard - Third Edition. In Book Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals; approved standard - Third Edition. City: CLSI; 2008. (Editor ed.^eds.)
• [27]Ewers C, Grobbel M, Stamm I, Kopp PA, Diehl I, Semmler T, Fruth A, Beutlich J, Guerra B, Wieler LH, Guenther S: Emergence of human pandemic O25:H4-ST131 CTX-M-15 extended-spectrum-beta-lactamase-producing Escherichia coli among companion animals. J Antimicrob Chemother 2010, 65:651-660.
• [28]Muller D, Greune L, Heusipp G, Karch H, Fruth A, Tschape H, Schmidt MA: Identification of unconventional intestinal pathogenic Escherichia coli isolates expressing intermediate virulence factor profiles by using a novel single-step multiplex PCR. Appl Environ Microbiol 2007, 73:3380-3390.
• [29]Schmidt H, Knop C, Franke S, Aleksic S, Heesemann J, Karch H: Development of PCR for screening of enteroaggregative Escherichia coli. J Clin Microbiol 1995, 33:701-705.
• [30]Toma C, Lu Y, Higa N, Nakasone N, Chinen I, Baschkier A, Rivas M, Iwanaga M: Multiplex PCR assay for identification of human diarrheagenic Escherichia coli. J Clin Microbiol 2003, 41:2669-2671.
• [31]Yamamoto T, Nakazawa M: Detection and sequences of the enteroaggregative Escherichia coli heat-stable enterotoxin 1 gene in enterotoxigenic E. coli strains isolated from piglets and calves with diarrhea. J Clin Microbiol 1997, 35:223-227.
• [32]Wirth T, Falush D, Lan R, Colles F, Mensa P, Wieler LH, Karch H, Reeves PR, Maiden MC, Ochman H, Achtman M: Sex and virulence in Escherichia coli: an evolutionary perspective. Molecular microbiology 2006, 60:1136-1151.
• [33]Okeke IN, Lamikanra A, Czeczulin J, Dubovsky F, Kaper JB, Nataro JP: Heterogeneous virulence of enteroaggregative Escherichia coli strains isolated from children in Southwest Nigeria. J Infect Dis 2000, 181:252-260.