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Applied and Environmental Microbiology, August 2006, p. 5359-5366, Vol. 72, No. 8
0099-2240/06/$08.00+0     doi:10.1128/AEM.00099-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Application of Bacteriophages To Control Intestinal Escherichia coli O157:H7 Levels in Ruminants

Haiqing Sheng, Hannah J. Knecht, Indira T. Kudva,{dagger} and Carolyn J. Hovde*

University of Idaho, Department of Microbiology, Molecular Biology, and Biochemistry, Moscow, Idaho 83844-3052

Received 13 January 2006/ Accepted 11 June 2006

A previously characterized O157-specific lytic bacteriophage KH1 and a newly isolated phage designated SH1 were tested, alone or in combination, for reducing intestinal Escherichia coli O157:H7 in animals. Oral treatment with phage KH1 did not reduce the intestinal E. coli O157:H7 in sheep. Phage SH1 formed clear and relatively larger plaques on lawns of all 12 E. coli O157:H7 isolates tested and had a broader host range than phage KH1, lysing O55:H6 and 18 of 120 non-O157 E. coli isolates tested. In vitro, mucin or bovine mucus did not inhibit bacterial lysis by phage SH1 or KH1. A phage treatment protocol was optimized using a mouse model of E. coli O157:H7 intestinal carriage. Oral treatment with SH1 or a mixture of SH1 and KH1 at phage/bacterium ratios ≥102 terminated the presence of fecal E. coli O157:H7 within 2 to 6 days after phage treatment. Untreated control mice remained culture positive for >10 days. To optimize bacterial carriage and phage delivery in cattle, E. coli O157:H7 was applied rectally to Holstein steers 7 days before the administration of 1010 PFU SH1 and KH1. Phages were applied directly to the rectoanal junction mucosa at phage/bacterium ratios calculated to be ≥102. In addition, phages were maintained at 106 PFU/ml in the drinking water of the phage treatment group. This phage therapy reduced the average number of E. coli O157:H7 CFU among phage-treated steers compared to control steers (P < 0.05); however, it did not eliminate the bacteria from the majority of steers.

* Corresponding author. Mailing address: Department of Microbiology, Molecular Biology, and Biochemistry, University of Idaho, Moscow, ID 83844-3052. Phone: (208) 885-5906. Fax: (208) 885-6518. E-mail: cbohach{at}uidaho.edu.

{dagger} Present address: Division of Infectious Diseases, Massachusetts General Hospital, Boston, MA 02114.

Applied and Environmental Microbiology, August 2006, p. 5359-5366, Vol. 72, No. 8
0099-2240/06/$08.00+0     doi:10.1128/AEM.00099-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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