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Applied and Environmental Microbiology, July 2008, p. 4070-4078, Vol. 74, No. 13
0099-2240/08/$08.00+0     doi:10.1128/AEM.00428-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Isolation and Characterization of Bacteriophages Infecting the Fish Pathogen Flavobacterium psychrophilum{triangledown}

Anne Rønnest Stenholm,1,2 Inger Dalsgaard,2 and Mathias Middelboe1*

Marine Biological Laboratory, University of Copenhagen, Strandpromenaden 5, 3000 Helsingør, Denmark,1 National Institute of Aquatic Resources, Fish Disease Laboratory, Technical University of Denmark, Stigbøjlen 4, 1870 Frederiksberg C, Denmark2

Received 21 February 2008/ Accepted 5 May 2008

Flavobacterium psychrophilum is a serious pathogen in trout aquaculture, responsible for the diseases rainbow trout fry syndrome (RTFS) and cold water disease (CWD). Bacteriophage control of F. psychrophilum may constitute a realistic approach in the treatment of these diseases; however, a detailed understanding of the phage-host interactions is needed to evaluate the potential of F. psychrophilum bacteriophages for that purpose. Twenty-two F. psychrophilum phages from Danish rainbow trout farms were isolated and characterized. The phage genome sizes differed considerably and fell into three major size classes (8.5 to 12 kb, 48 kb, and 90 kb). The phage host ranges comprised from 5 to 23 of the 28 tested F. psychrophilum strains, and 18 of the phage isolates showed unique host ranges. Each bacterial strain had a unique pattern of susceptibility to the 22 phages, and individual strains also showed large variations (up to 107-fold differences) in susceptibility to specific phages. Phage burst size (7 to 162 phages infected cell–1) and latency period (4 to 6 h) also showed pronounced differences both between phages and, for a specific phage, between host strains. In general, the characterization documented the presence of diverse F. psychrophilum phage communities in Danish trout farms, with highly variable patterns of infectivity. The discovery and characterization of broad-host-range phages with strong lytic potential against numerous pathogenic F. psychrophilum host strains thus provided the foundation for future exploration of the potential of phages in the treatment of RTFS and CWD.

* Corresponding author. Mailing address: Marine Biological Laboratory, University of Copenhagen, Strandpromenaden 5, 3000 Helsingør, Denmark. Phone: 45 3532 1991. Fax: 45 3532 1951. E-mail: MMiddelboe{at}bio.ku.dk

{triangledown} Published ahead of print on 9 May 2008.

Applied and Environmental Microbiology, July 2008, p. 4070-4078, Vol. 74, No. 13
0099-2240/08/$08.00+0     doi:10.1128/AEM.00428-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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