Rapid Diagnosis of Red Spot Disease in Cultured Eels by Direct Immunofluorescence (IF)

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  • 免疫けい光直接法による養殖ウナギ赤点病の迅速診断
  • 免疫螢光直接法による養殖ウナギ赤点病の迅速診断
  • メンエキ ケイコウ チョクセツホウ ニヨル ヨウショク ウナギ セキテンビョウ

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Red spot disease of cultured eels, caused by Pseudomonas anguilliseptica, has been occurring in Japan since the early summer of 1971. Direct immunofluorescence (IF) was examined for rapid diagnosis of the disease in this study.<br> Antiserum was obtained from rabbit immunized with formalin-killed organisms (strain ST-1), The titer was 1:2048 by the agglutination test. IgG fraction was purified from the antiserum by ammonium sulfate precipitation and conjugated with fluorescein isothiocyanate. Free dye was removed by Sephadex gel filtration and the conjugate was purified by DEAE-cellulose column chromatography.<br> The F/P molar ratio of the conjugate was determined as 1.2. The conjugate had a staining titer of 1:32 and no cross-reaction was observed when Vibrio fisheri, Edwardsiella tarda, Streptococcus sp, and Ps. fluorescens were stained, but slight cross-reaction was seen in low dilution with Ps. putida, Ps. aeruginosa, Aeromonas sp. and Vibrio anguillarum.<br> The specific fluorescence in ST-1 cells was observed when acetone-fixed impression preparalion or frozen section from artificially infected eels was stained. ST-1 cell was more readily detected by IF than cultivation on nutrient agar. The specific fluorescence could be observed at least sixteen days at 37°C.<br> Although the conjugate was not evaluated on natural epizootics, the above data suggest that we could employ direct IF on improssion preparations for the rapid and reliable diagnosis of this disease.

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