Variation in Stress Resistance Patterns among stx Genotypes and Genetic Lineages of Shiga Toxin-Producing Escherichia coli O157

ABSTRACT To evaluate the relationship between bacterial genotypes and stress resistance patterns, we exposed 57 strains of Shiga toxin-producing Escherichia coli (STEC) O157 to acid, freeze-thaw, heat, osmotic, oxidative, and starvation stresses. Inactivation rates were calculated in each assay and subjected to univariate and multivariate analyses, including principal component analysis (PCA) and cluster analysis. The stx genotype was determined for each strain as was the lineage-specific polymorphism assay (LSPA6) genotype. In univariate analyses, strains of the stx 1 stx 2 genotype showed greater resistance to heat than strains of the stx 1 stx 2c genotype; moreover, strains of the stx 1 stx 2 genotype showed greater resistance to starvation than strains of the stx 2 or stx 2c genotypes. LSPA6 lineage I (LI) strains showed greater resistance to heat and starvation than LSPA6 lineage II (LII) strains. PCA revealed a general trend that a strain with greater resistance to one type of stress tended to have greater resistance to other types of stresses. In cluster analysis, STEC O157 strains were grouped into stress-resistant, stress-sensitive, and intermediate clusters. In stx genotypes, all strains of the stx 1 stx 2 genotype were grouped with the stress-resistant cluster, whereas 72.7% (8/11) of strains of the stx 1 stx 2c genotype grouped with the stress-sensitive cluster. In LI strains, 77.8% (14/18) of the strains were grouped with the stress-resistant cluster, whereas 64.7% (11/17) of LII strains were grouped with the stress-sensitive cluster. These results indicate that the genotypes of STEC O157 that are frequently associated with human illness, i.e., LI or the stx 1 stx 2 genotype, have greater multiple stress resistance than do strains of other genotypes.

[1]  T. Whittam,et al.  Variation in virulence among clades of Escherichia coli O157:H7 associated with disease outbreaks , 2008, Proceedings of the National Academy of Sciences.

[2]  Elaine R. Mardis,et al.  A precise reconstruction of the emergence and constrained radiations of Escherichia coli O157 portrayed by backbone concatenomic analysis , 2009, Proceedings of the National Academy of Sciences.

[3]  Ali S. Hadi,et al.  Finding Groups in Data: An Introduction to Chster Analysis , 1991 .

[4]  R. Mckellar,et al.  Growth and survival of various strains of enterohemorrhagic Escherichia coli in hydrochloric and acetic acid. , 1999, Journal of food protection.

[5]  William N. Venables,et al.  Modern Applied Statistics with S , 2010 .

[6]  T. Whittam,et al.  Increased Adherence and Expression of Virulence Genes in a Lineage of Escherichia coli O157:H7 Commonly Associated with Human Infections , 2010, PloS one.

[7]  R. Mandrell,et al.  Escherichia coli O157:H7 strains isolated from environmental sources differ significantly in acetic acid resistance compared with human outbreak strains. , 2009, Journal of food protection.

[8]  Paul J. Watson,et al.  Statistics for Veterinary and Animal Science , 1999 .

[9]  A. Benson,et al.  Identification of Common Subpopulations of Non-Sorbitol-Fermenting, β-Glucuronidase-Negative Escherichia coli O157:H7 from Bovine Production Environments and Human Clinical Samples , 2004, Applied and Environmental Microbiology.

[10]  D. Collet Modelling Survival Data in Medical Research , 2004 .

[11]  G. Storz,et al.  Bacterial stress responses. , 2011 .

[12]  R Core Team,et al.  R: A language and environment for statistical computing. , 2014 .

[13]  Sang Yup Lee,et al.  The Escherichia coli Proteome: Past, Present, and Future Prospects , 2006, Microbiology and Molecular Biology Reviews.

[14]  James M. Jay,et al.  Modern food microbiology , 1970 .

[15]  F. Dziva,et al.  Options for the control of enterohaemorrhagic Escherichia coli in ruminants. , 2002, Microbiology.

[16]  J. Bono,et al.  Greater Diversity of Shiga Toxin-Encoding Bacteriophage Insertion Sites among Escherichia coli O157:H7 Isolates from Cattle than in Those from Humans , 2006, Applied and Environmental Microbiology.

[17]  A. Fruth,et al.  Enterohemorrhagic Escherichia coli. , 2001, Contributions to microbiology.

[18]  Eduardo N. Taboada,et al.  In silico genomic analyses reveal three distinct lineages of Escherichia coli O157:H7, one of which is associated with hyper-virulence , 2009, BMC Genomics.

[19]  K. Horikawa,et al.  Biased distribution of IS629 among strains in different lineages of enterohemorrhagic Escherichia coli serovar O157. , 2011, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[20]  A. Benson,et al.  Influence of animal origin and lineage on survival of Escherichia coli O157:H7 strains in strong and weak acid challenges. , 2004, Journal of food protection.

[21]  M Peleg,et al.  Reinterpretation of microbial survival curves. , 1998, Critical reviews in food science and nutrition.

[22]  M. Rowe,et al.  Effect of nutrient starvation on the resistance of Escherichia coli O157:H7 to subsequent heat stress. , 2000, Journal of food protection.

[23]  Brian D. Ripley,et al.  Modern applied statistics with S, 4th Edition , 2002, Statistics and computing.

[24]  S. Iyoda,et al.  Multivariate Analyses Revealed Distinctive Features Differentiating Human and Cattle Isolates of Shiga Toxin-Producing Escherichia coli O157 in Japan , 2011, Journal of Clinical Microbiology.

[25]  A. Benson,et al.  Genotypic Characterization and Prevalence of Virulence Factors among Canadian Escherichia coli O157:H7 Strains , 2008, Applied and Environmental Microbiology.

[26]  H. Vogel,et al.  Acetylornithinase of Escherichia coli: partial purification and some properties. , 1956, The Journal of biological chemistry.

[27]  M. Drake,et al.  Stress Response of Escherichia coli , 2006 .

[28]  S. Dowd,et al.  Microarray based comparison of two Escherichia coli O157:H7 lineages , 2006, BMC Microbiology.

[29]  M. Drake,et al.  Acid stress, starvation, and cold stress affect poststress behavior of Escherichia coli O157:H7 and nonpathogenic Escherichia coli. , 2001, Journal of food protection.

[30]  Teresa M. Bergholz,et al.  Salt stress phenotypes in Listeria monocytogenes vary by genetic lineage and temperature. , 2010, Foodborne pathogens and disease.

[31]  Takashi Yura,et al.  Convergence of Molecular, Modeling, and Systems Approaches for an Understanding of the Escherichia coli Heat Shock Response , 2008, Microbiology and Molecular Biology Reviews.

[32]  Olaf Berke,et al.  Statistics for Veterinary and Animal Science, 2nd ed. , 2007 .

[33]  T. Zotta,et al.  Diversity of stress responses in dairy thermophilic streptococci. , 2008, International journal of food microbiology.

[34]  G. Schoolnik,et al.  Genomic and Phenotypic Diversity of Coastal Vibrio cholerae Strains Is Linked to Environmental Factors , 2007, Applied and Environmental Microbiology.

[35]  S. Torriani,et al.  Diversity of stress tolerance in Lactobacillus plantarum, Lactobacillus pentosus and Lactobacillus paraplantarum: A multivariate screening study. , 2010, International journal of food microbiology.

[36]  J. Jacobs,et al.  Influence of Environmental Gradients on the Abundance and Distribution of Mycobacterium spp. in a Coastal Lagoon Estuary , 2009, Applied and Environmental Microbiology.

[37]  M. Miyahara,et al.  Loss of O157 O Antigenicity of Verotoxin-ProducingEscherichia coli O157:H7 Surviving under Starvation Conditions , 2001, Applied and Environmental Microbiology.

[38]  T. Robinson,et al.  Variation in Resistance of Natural Isolates ofEscherichia coli O157 to High Hydrostatic Pressure, Mild Heat, and Other Stresses , 1999, Applied and Environmental Microbiology.

[39]  E. Dudley,et al.  Escherichia coli O157:H7 of genotype lineage-specific polymorphism assay 211111 and clade 8 are common clinical isolates within Pennsylvania. , 2011, Foodborne pathogens and disease.

[40]  Chad R. Laing,et al.  Lineage and Host Source Are Both Correlated with Levels of Shiga Toxin 2 Production by Escherichia coli O157:H7 Strains , 2009, Applied and Environmental Microbiology.

[41]  P. McClure,et al.  Survival of Escherichia coli in foods , 2000, Symposium series.

[42]  M. Kleerebezem,et al.  Phenotypic and genomic diversity of Lactobacillus plantarum strains isolated from various environmental niches. , 2010, Environmental microbiology.

[43]  David Collett Modelling Survival Data in Medical Research , 1994 .

[44]  C. Gyles Shiga toxin-producing Escherichia coli: an overview. , 2007, Journal of animal science.

[45]  K. Ouchi,et al.  Principal-Component Analysis of the Characteristics Desirable in Baker's Yeasts , 1989, Applied and environmental microbiology.

[46]  T. Whittam,et al.  Differential Expression of Virulence and Stress Fitness Genes between Escherichia coli O157:H7 Strains with Clinical or Bovine-Biased Genotypes , 2009, Applied and Environmental Microbiology.

[47]  L. Brandt,et al.  Escherichia coli 0157 : H 7 Infection in Humans , 2009 .

[48]  J. Lejeune,et al.  Association of prophage antiterminator Q alleles and susceptibility to food-processing treatments applied to Escherichia coli O157 in laboratory media. , 2007, Journal of Food Protection.

[49]  A. Benson,et al.  Octamer-based genome scanning distinguishes a unique subpopulation of Escherichia coli O157:H7 strains in cattle. , 1999, Proceedings of the National Academy of Sciences of the United States of America.