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Comparison of Enrichment Broths for Supporting Growth of Shiga Toxin-Producing Escherichia coli

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Abstract

Detection of Shiga toxin-producing Escherichia coli (STEC) in complex sample matrices remains challenging. In an attempt to improve detection, nonselective and selective enrichment broths were compared as follows: (1) trypticase soy broth (TSB) was compared with TSB plus novobiocin, vancomycin, rifampicin, bile salts, and potassium tellurite (TSB-NVRBT) for supporting growth of STEC in pure culture; (2) E. coli broth (EC), TSB, and TSB plus bile salts (mTSB) were compared for enrichment of STEC O26, O45, O103, O104, O111, O121, O145, and O157 (STEC-8) in inoculated cattle fecal samples; (3) EC, TSB, and mTSB were compared for the detection of STEC-8 in inoculated cattle fecal samples. Fecal samples were inoculated with wild-type STEC-8 or nalidixic acid- or rifampicin-resistant derivatives of the same strains at 100, 1000, or 10,000 colony-forming units per gram (CFU/g) of feces. In pure culture, the mean STEC CFU/mL following enrichment in TSB was 1.17 log10 greater than that in TSB-NVRBT (P < 0.05). In inoculated fecal samples, EC enrichment yielded growth of STEC-8 (6.42 log10 CFU/g) that was significantly greater than in TSB (6.23 log10 CFU/g; P < 0.05), and numerically but not significantly greater than in mTSB (6.37 log10 CFU/g; P = 0.60). Wild-type STEC strains were detected in 43.8 % (21/48) of the samples enriched in EC and mTSB compared to 27.1 % (13/48) of the samples enriched in TSB (P = 0.15). Overall, STEC grew significantly better when enriched in EC compared to TSB. Modification of TSB by the addition of bile salts improved the growth and detection of STEC compared to TSB alone.

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References

  1. Barkocy-Gallagher GA, Edwards KK, Nou X, Bosilevac JM, Arthur TM, Shackelford SD, Koohmaraie M (2005) Methods for recovering Escherichia coli O157:H7 from cattle fecal, hide and carcass samples: sensitivity and improvements. J Food Prot 68:2264–2268

    PubMed  Google Scholar 

  2. Baylis CL (2008) Growth of pure cultures of verocytotoxin-producing Escherichia coli in a range of enrichment media. J Appl Microbiol 105:1259–1265

    Article  CAS  PubMed  Google Scholar 

  3. Bettelheim KA (2007) The non-O157 Shiga-toxigenic (verocytotoxigenic) Escherichia coli; under-rated pathogens. Crit Rev Microbiol 33:67–87

    Article  PubMed  Google Scholar 

  4. Brooks JT, Sowers EG, Wells JG, Greene KD, Griffin PM, Hoekstra RM, Strockbine NA (2005) Non-O157 Shiga toxin-producing Escherichia coli infections in the United States, 1983–2002. J Infect Dis 92:1422–1429

    Article  Google Scholar 

  5. Bucholz U, Bernard H, Werber D, Böhmer MM, Remschmidt C, Wilking H, Deleré Y, an der Heiden M, Adlhoch C, Dreesman J, Ehlers J, Ethelberg S, Faber M, Frank C, Fricke G, Greiner M, Höhle M, Ivarsson S, Jark U, Kirchner M, Koch J, Krause G, Luber P, Rosner B, Stark K, Kühne M (2011) German outbreak of Escherichia coli O104:H4 associated with sprouts. N Engl J Med 365:1763–1770

    Article  Google Scholar 

  6. Fegan N, Higgs G, Vanderlinde P, Desmarchelier P (2004) Enumeration of Escherichia coli O157 in cattle faeces using most probable number technique and automated immunomagnetic separation. Lett Appl Microbiol 38:56–59

    Article  CAS  PubMed  Google Scholar 

  7. Fukushima H, Gomyoda M (1999) An effective, rapid and simple method for isolation of Shiga toxin-producing Escherichia coli O26, O111 and O157 from faeces and food samples. Zentralbl Bakteriol 289:415–428

    Article  CAS  PubMed  Google Scholar 

  8. Hussein HS, Bollinger LM (2008) Influence of selective media on successful detection of Shiga toxin-producing Escherichia coli in food, fecal, and environmental samples. Foodborne Pathog Dis 5:227–244

    Article  CAS  PubMed  Google Scholar 

  9. Hussein HS, Bollinger LM, Hall MR (2008) Growth and enrichment medium for detection and isolation of Shiga toxin-producing Escherichia coli in cattle feces. J Food Prot 71:927–933

    PubMed  Google Scholar 

  10. Kanki M, Seto K, Harada T, Yonogi S, Kumeda Y (2011) Comparison of four enrichment broths for the detection of non-O157 Shiga toxin-producing Escherichia coli O91, O103, O111, O119, O121, O145 and O165 from pure culture and food samples. Lett Appl Microbiol 53:167–173

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  12. Maturin L, Peeler JT (2001) Aerobic plate count. In: Merker RI (ed) Bacteriological analytical manual, 8th ed. revision A U.S. Food and Drug Administration, College Park, MD. http://www.fda.gov/food/foodscienceresearch/laboratorymethods/ucm2006949.htm. Accessed 25 Feb 2013

  13. Paddock Z, Shi X, Bai J, Nagaraja TG (2012) Applicability of a multiplex PCR to detect O26, O45, O103, O111, O121, O145, and O157 serogroups of Escherichia coli in cattle feces. Vet Microbiol 156:381–388

    Article  CAS  PubMed  Google Scholar 

  14. Possé B, De Zutter L, Heyndrickx M, Herman L (2008) Novel differential and confirmation plating media for Shiga toxin-producing Escherichia coli serotypes O26, O103, O111, O145 and sorbitol-positive and -negative O157. FEMS Microbiol Lett 282:124–131

    Article  PubMed  Google Scholar 

  15. Possé B, De Zutter L, Heyndrickx M, Herman L (2008) Quantitative isolation efficiency of O26, O103, O111, O145 and O157 STEC serotypes from artificially contaminated food and cattle faeces samples using a new isolation protocol. J Appl Microbiol 105:227–235

    Article  PubMed  Google Scholar 

  16. Sánchez S, Garciá-Sánchez A, Martínez R, Blanco J, Blanco JE, Blanco M, Dahbi G, Mora A, Hermoso de Mendoza J, Alonso JM, Rey J (2009) Detection and characterisation of Shiga toxin-producing Escherichia coli other than Escherichia coli O157:H7 in wild ruminants. Vet J 180:384–388

    Article  PubMed  Google Scholar 

  17. Sanderson MW, Gay JM, Hancock DD, Gay CC, Fox LK, Besser TE (1995) Sensitivity of bacteriologic culture for detection of Escherichia coli O157:H7 in bovine feces. J Clin Microbiol 33:2616–2619

    CAS  PubMed Central  PubMed  Google Scholar 

  18. Tilden J Jr, Young W, McNamara AM, Custer C, Boesel B, Lambert-Fair MA, Majkowski J, Vugia D, Werner SB, Hollingsworth Morris JG Jr (1996) A new route of transmission for Escherichia coli: infection from dry fermented salami. Am J Public Health 86:1142–1150

    Article  PubMed Central  PubMed  Google Scholar 

  19. Verstraete K, De Zutter L, Messens W, Herman L, Heyndrickx M, De Reu K (2010) Effect of the enrichment time and immunomagnetic separation on the detection of Shiga toxin-producing Escherichia coli O26, O103, O111, O145 and sorbitol positive O157 from artificially inoculated cattle faeces. Vet Microbiol 145:106–112

    Article  PubMed  Google Scholar 

  20. Vimont A, Vernozy-Rozand C, Delignette-Muller ML (2006) Isolation of E. coli O157:H7 and non-O157 STEC in different matrices: review of the most commonly used enrichment protocols. Lett Appl Microbiol 42:102–108

    Article  CAS  PubMed  Google Scholar 

  21. Vimont A, Vernozy-Rozand C, Montet MP, Bavai C, Fremaux B, Delignette-Muller ML (2007) Growth of Shiga-toxin producing Escherichia coli (STEC) and bovine feces background microflora in various enrichment protocols. Vet Microbiol 123:274–281

    Article  CAS  PubMed  Google Scholar 

  22. Wang F, Yang Q, Kase JA, Meng J, Clotilde LM, Lin A, Ge B (2013) Current trends in detecting non-O157 Shiga toxin-producing Escherichia coli in food. Foodborne Pathog Dis 10:665–677

    Article  PubMed  Google Scholar 

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Acknowledgments

This project was supported by Agriculture and Food Research Initiative Competitive Grant No. 2012-68003-30155 from the USDA National Institute of Food and Agriculture. The authors thank Drs. Shannon Manning, David Renter, and John Luchansky for sharing strains.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln, 1880 N. 42nd St., Lincoln, NE, 68583-0905, USA

    Zachary R. Stromberg, Gentry L. Lewis & Rodney A. Moxley

  2. Department of Statistics, University of Nebraska-Lincoln, Lincoln, NE, USA

    David B. Marx

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  1. Zachary R. Stromberg
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  2. Gentry L. Lewis
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  3. David B. Marx
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  4. Rodney A. Moxley
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Correspondence to Rodney A. Moxley.

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Stromberg, Z.R., Lewis, G.L., Marx, D.B. et al. Comparison of Enrichment Broths for Supporting Growth of Shiga Toxin-Producing Escherichia coli . Curr Microbiol 71, 214–219 (2015). https://doi.org/10.1007/s00284-015-0824-8

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  • DOI: https://doi.org/10.1007/s00284-015-0824-8

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