In January 2003 the leading 200 beef industry companies held an unprecedented meeting to develop a """"""""unified battle plan"""""""" to eliminate E. coli O157 from their industry using methods based in sound science. This important food-borne pathogen causes >70,000 illnesses yearly in the U.S., >75% of them directly linked to livestock; at present 28% of U.S. cattle carry O157, though they show no symptoms. Preliminary experiments supported by our current NIH/AREA grant suggest that, with targeted research and development, bacteriophages could play a significant role in protecting our food supply. We isolated CEV1, the T4-like phage we have used most, from a flock of sheep found to be highly resistant to gut colonization by O157 during probiotic studies at the Southern Plains USDA/ARS Research Center, our collaborators on this project. We have since isolated promising phages from two other flocks of sheep. The long-term objectives of this proposal are to (1) isolate and characterize further phages capable of infecting E. coli O157:H7, (2) investigate the dynamics and physiology of replication of those phages under conditions relevant to the natural environment and to potential therapeutic applications, i. e. during anaerobic respirative and fermentative growth of the host, and select the most promising phages and multiphage cocktails; and (3) use appropriately-designed mixed-culture fermenters and experiments in sheep to assess the potential for using these phages to reduce the O157:H7 load in the guts of livestock and thus in the foodchain. Such experiments are very well suited to work by our undergraduate students, helping them develop a firm foundation in microbial ecology and physiology, molecular biology, general microbial and anaerobic techniques, and experimental design, along with the satisfaction of contributing significant new research results. In preliminary studies, O157:H7 was successfully eradicated from gut models by CEV1 treatment in 12 days and O157 levels were markedly reduced in sheep in two days by CEV1 and CEV2.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Academic Research Enhancement Awards (AREA) (R15)
Project #
2R15GM063637-02
Application #
6754735
Study Section
Bacteriology and Mycology Subcommittee 2 (BM)
Program Officer
Anderson, James J
Project Start
2001-08-01
Project End
2006-12-31
Budget Start
2004-04-01
Budget End
2006-12-31
Support Year
2
Fiscal Year
2004
Total Cost
$205,724
Indirect Cost
Name
Evergreen State College
Department
Miscellaneous
Type
Schools of Arts and Sciences
DUNS #
069579977
City
Olympia
State
WA
Country
United States
Zip Code
98505
Bryan, Daniel; El-Shibiny, Ayman; Hobbs, Zack et al. (2016) Bacteriophage T4 Infection of Stationary Phase E. coli: Life after Log from a Phage Perspective. Front Microbiol 7:1391
Adriaenssens, Evelien M; Ackermann, Hans-Wolfgang; Anany, Hany et al. (2012) A suggested new bacteriophage genus: ""Viunalikevirus"". Arch Virol 157:2035-46
Kutter, Elizabeth M; Skutt-Kakaria, Kyobi; Blasdel, Bob et al. (2011) Characterization of a ViI-like phage specific to Escherichia coli O157:H7. Virol J 8:430
Raya, Raul R; Varey, Peter; Oot, Rebecca A et al. (2006) Isolation and characterization of a new T-even bacteriophage, CEV1, and determination of its potential to reduce Escherichia coli O157:H7 levels in sheep. Appl Environ Microbiol 72:6405-10