title: Complete genome sequence analysis of two Pseudomonas plecoglossicida phages, potential therapeutic agents.
abstract: Pseudomonas plecoglossicida is a lethal pathogen of ayu (Plecoglossus altivelis) in Japan and is responsible for substantial economic costs to ayu culture. Previously, we demonstrated the efficacy of phage therapy against P. plecoglossicida infection using two lytic phages (PPpW-3 and PPpW-4) (S. C. Park, I. Shimamura, M. Fukunaga, K. Mori, and T. Nakai, Appl Environ Microbiol 66:1416-1422, 2000, http://dx.doi.org/10.1128/AEM.66.4.1416-1422.2000; S. C. Park and T. Nakai, Dis Aquat Org 53:33-39, 2003, http://dx.doi.org/10.3354/dao053033). In the present study, the complete genome sequences of these therapeutic P. plecoglossicida phages were determined and analyzed for deleterious factors as therapeutic agents. The genome of PPpW-3 (myovirus) consisted of 43,564 bp with a GC content of 61.1% and 66 predicted open reading frames (ORFs). Approximately half of the genes were similar to the genes of the Escherichia coli phage vB_EcoM_ECO1230-10 (myovirus). The genome of PPpW-4 (podovirus) consisted of 41,386 bp with a GC content of 56.8% and 50 predicted ORFs. More than 70% of the genes were similar to the genes of Pseudomonas fluorescens phage ϕIBB-PF7A and Pseudomonas putida phage ϕ15 (podoviruses). The whole-genome analysis revealed that no known virulence genes were present in PPpW-3 and PPpW-4. An integrase gene was found in PPpW-3, but other factors used for lysogeny were not confirmed. The PCR detection of phage genes in phage-resistant variants provided no evidence of lysogenic activity in PPpW-3 and PPpW-4. We conclude that these two lytic phages qualify as therapeutic agents.
authors.affiliation: Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima, Japan ykawato@affrc.go.jp.
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Kawato
authors.affiliation: Research Center for Aquatic Genomics, National Research Institute of Fisheries Science, Fisheries Research Agency, Yokohama, Japan.
authors.forename: Motoshige
authors.initials: M
authors.lastname: Yasuike
authors.affiliation: Research Center for Aquatic Genomics, National Research Institute of Fisheries Science, Fisheries Research Agency, Yokohama, Japan.
authors.forename: Yoji
authors.initials: Y
authors.lastname: Nakamura
authors.affiliation: Research Center for Fisheries Oceanography and Marine Ecosystem, National Research Institute of Fisheries Science, Fisheries Research Agency, Yokohama, Japan.
authors.forename: Yuya
authors.initials: Y
authors.lastname: Shigenobu
authors.affiliation: Research Center for Aquatic Genomics, National Research Institute of Fisheries Science, Fisheries Research Agency, Yokohama, Japan.
authors.forename: Atushi
authors.initials: A
authors.lastname: Fujiwara
authors.affiliation: Research Center for Aquatic Genomics, National Research Institute of Fisheries Science, Fisheries Research Agency, Yokohama, Japan.
authors.forename: Motohiko
authors.initials: M
authors.lastname: Sano
authors.affiliation: Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima, Japan.
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Nakai
mesh_terms: D000818:Animals; D001482:Base Composition; D004279:DNA, Viral / Q000737:chemistry* / Q000235:genetics*; D016679:Genome, Viral; D007564:Japan; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D036441:Osmeriformes / Q000382:microbiology; D011549:Pseudomonas / Q000821:virology; D017105:Pseudomonas Phages / Q000235:genetics*; D017422:Sequence Analysis, DNA; D017385:Sequence Homology
chemical_list: D004279:DNA, Viral