Quantitative proteomics (iTRAQ)-based analysis of the O157 anaerobic proteome expressed in uRF with all normal rumen flora was performed to more closely determine O157 protein expression in the bovine rumen. The cumulative results of all RF-preparation analysis suggested that rumen specific protein expression enables O157 to adapt to this hostile environment and successfully transit to its colonization sites in the bovine GIT. To further verify our conclusions, we are evaluating the O157 proteomic-profile as expressed in vivo in a rumen-fistulated cow, and confirming the role of a subset of these

‘adaptive’ proteins in O157 survival. Acknowledgements Technical support provided by Bryan Wheeler, Deb Hinrichsen (NVSL) and Laurie Evans (NVSL)

in collection DZNeP purchase & filtration of rumen fluid; Deb Lebo and Sam Humphrey in analyzing VFAs; Duane Zimmerman for assisting with iTRAQ labeling and Paul Amundson’s group of animal caretakers for assisting in rumen fluid collection is acknowledged with appreciation. Bottom-up proteomics was done at the Proteomics Division, ICBR, University of Florida, Gainesville, FL. We thank Dr. Manohar John, Dr. Thomas Casey and Dr. John Bannantine for their insightful PU-H71 concentration review of this manuscript. Disclaimer Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture. USDA is an equal opportunity provider and MM-102 cost employer. Electronic supplementary material Additional file 1: Table S1: Bottom-up Proteomics Dataset. (XLS 890 KB) Additional file 2: Table S2: iTRAQ Proteomics Dataset. (XLS 166 KB) References 1. Scallan E, Hoekstra RM, Angulo FJ, Tauxe RV, Widdowson M, Roy SL, Jones JL, Griffin PM: Foodborne illness acquired in the United States – Major pathogens. J Animal Sci 2011, 17:7–15. 2. Vital signs: Incidence and trends of infection with pathogens transmitted commonly Etomidate through food — Foodborne

diseases active surveillance network, 10 U.S. Sites, 1996–2010. MMWR 2011, 60:749–755. 3. CDC: Preliminary FoodNet data on the incidence of infection with pathogens transmitted commonly through food–10 sites, United States, 2004. MMWR 2005, 54:352–356. 4. Griffin PM, Ostroff SM, Tauxe RV, Greene KD, Wells JG, Lewis JH, Blake PA: Illnesses associated with Escherichia coli 0157:H7 infections A broad clinical spectrum. Ann Intern Med 1998, 109:705–712.CrossRef 5. Kaper JB, O’Brien AD: Escherichia coli O157:H7 and other Shiga Toxin-Producing E. coli Strains. Washington, D.C: ASM Press; 1998. 6. Wolin MJ: Volatile fatty acids and the inhibition of Escherichia coli growth by rumen fluid. Appl Microbiol 1969, 17:83–87.PubMedCentralPubMed 7. Schneider IC, Ames ML, Rasmussen MA, Reilly PJ: Fermentation of cottonseed and other feedstuffs in cattle rumen fluid. J Agric Food Chem 2002, 50:2267–2273.PubMedCrossRef 8.