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Applied and Environmental Microbiology, April 2000, p. 1416-1422, Vol. 66, No. 4
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Isolation of Bacteriophages Specific to a Fish Pathogen, Pseudomonas plecoglossicida, as a Candidate for Disease Control

Se Chang Park,1 Ichiro Shimamura,2 Minoru Fukunaga,2 Koh-Ichiro Mori,3 and Toshihiro Nakai1,*

Fish Pathology Laboratory, Faculty of Applied Biological Science, Hiroshima University, Higashi-hiroshima, Hiroshima,1 Tokushima Prefectural Fisheries Experimental Station, Hiwasa, Tokushima,2 and Kamiura Station, Japan Sea-Farming Association, Kamiura, Oita,3 Japan

Received 25 October 1999/Accepted 6 January 2000

Two types of bacteriophage specific to Pseudomonas plecoglossicida, the causative agent of bacterial hemorrhagic ascites disease in cultured ayu fish (Plecoglossus altivelis), were isolated from diseased ayu and the rearing pond water. One type of phage, which formed small plaques, was tentatively classified as a member of the family Myoviridae, and the other type, which formed large plaques, was classified as a member of the family Podoviridae. All 27 strains of P. plecoglossicida examined, which were isolated from diseased ayu from geographically different areas in 1991 to 1999, exhibited quite similar sensitivities to either type of phage. One strain of P. plecoglossicida was highly virulent for ayu, and the 50% lethal dose (LD50) when intramuscular injection was used was 101.2 CFU fish-1; in contrast, phage-resistant variants of this organism were less virulent (LD50, >104 CFU fish-1). Oral administration of phage-impregnated feed to ayu resulted in protection against experimental infection with P. plecoglossicida. After oral administration of P. plecoglossicida cells of this bacterium were always detected in the kidneys of control fish that did not receive the phage treatment, while the cells quickly disappeared from the phage-treated fish. Bacterial growth in freshwater was lower in the presence of phage, and the number of phage PFU increased rapidly. These results suggest that it may be possible to use phage to control the disease caused by P. plecoglossicida.

* Corresponding author. Mailing address: Applied Biological Science, Hiroshima University, Higashihiroshima 739-8528, Japan. Phone: 81-824-24-7947. Fax: 81-824-22-7059. E-mail: nakaitt{at}hiroshima-u.ac.jp.

Applied and Environmental Microbiology, April 2000, p. 1416-1422, Vol. 66, No. 4
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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