DAO 108:103-112 (2014)  -  DOI: https://doi.org/10.3354/dao02679

Antigenic proteins of Flavobacterium psychrophilum recognized by ayu Plecoglossus altivelis antisera

Goshi Kato1,2, Takamitsu Sakai1, Kyuma Suzuki3, Kenichi Yamaguchi4, Tomokazu Takano5, Tomomasa Matsuyama5, Chihaya Nakayasu1,* 

1Tamaki Laboratory, National Research Institute of Aquaculture, Fisheries Research Agency, Tamaki, Mie 519-0423, Japan
2The Japan Society for the Promotion of Science, 102-0083, Chiyoda, Tokyo, Japan
3Gunma Prefectural Fisheries Experimental Station, Maebashi, Gunma 371-0036, Japan
4Faculty of Fisheries, Nagasaki University, Nagasaki, Nagasaki 852-8521, Japan
5National Research Institute of Aquaculture, Fisheries Research Agency, Minami-Ise, Mie 516-0193, Japan
*Corresponding author:

ABSTRACT: Flavobacterium psychrophilum is the causative agent of bacterial coldwater disease (BCWD) in ayu Plecoglossus altivelis altivelis and is responsible for substantial economic losses in ayu culture in Japan. To develop effective vaccines for the disease, we identified antigenic proteins of F. psychrophilum using immunoglobulin from ayu that had recovered from BCWD. The whole protein extracted from the bacterium was separated using 2-dimensional polyacrylamide gel electrophoresis and was transferred to a polyvinylidene fluoride membrane. Subsequently, antigenic proteins of the bacterium were detected using western blotting and ayu antisera against F. psychrophilum. Each protein spot showing antigenicity was subjected to tandem mass spectrometry (MS/MS) analysis using a MALDI-QIT-TOF mass spectrometer. Protein identification based on the MS/MS data was performed using the genome database for F. psychrophilum JIP02/86, and the subcellular localization for each identified protein was predicted with web-based tools (LipoP and PSORTb). In total, 62 antigenic proteins were identified: of these, 46 were putative cytoplasmic proteins (e.g. elongation factor Tu and heat shock protein 90). The remaining 21 proteins were identified as putative membrane-bound or secreted proteins and are potential vaccine candidates. These proteins include OmpA, Omp 121, M13 family metallopeptidase, and M48 family metalloprotease.


KEY WORDS: Bacterial coldwater disease · BCWD · 2D-PAGE · MS/MS analysis · Vaccine · Anti-ayu immunoglobin MAb


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Cite this article as: Kato G, Sakai T, Suzuki K, Yamaguchi K, Takano T, Matsuyama T, Nakayasu C (2014) Antigenic proteins of Flavobacterium psychrophilum recognized by ayu Plecoglossus altivelis antisera. Dis Aquat Org 108:103-112. https://doi.org/10.3354/dao02679

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