To develop an accurate diagnosis of "new ulcer disease" in koi carp, we produced four monoclonal antibodies (mAbs) against a strain of atypical Aeromonas salmonicida isolated from koi carp. These mAbs did not cross-react with an isolate of atypical A. salmonicida from Japanese flounder and other pathogenic bacteria. Re-isolation from artificially infected koi carp was achieved by selecting the blue colonies on agar medium containing Coomassie brilliant blue, and some of the colonies were detected by immunofluorescent staining using the mAbs. These results suggested that the mAbs can distinguish atypical A. salmonicida from koi carp from resident aeromonads.
Infectious disease is one of the most serious problems in the aquaculture industry for ornamental or edible fish. This study attempted to develop a new device for preventing an aquatic bacterial disease, ulcer disease, caused by Aeromonas salmonicida (As), using “affinity silk”. Affinity silk is a silk protein-containing fibroin L-chain (FibL) fused to the single-chain variable fragment (scFv). It can be easily processed into different formats such as fibers, gels, sponges, or films. A transgenic silkworm that could express a cDNA construct containing FibL fused to an scFv derived from a monoclonal antibody (MAb) against As was successfully generated. An enzyme-linked immunosorbent assay was used to detect As by employing 96-well plates coated with scFv-conjugated affinity silk. As could be captured efficiently by glass wool coated with affinity silk in the column. Furthermore, the air-lift water filter equipped with the affinity silk-coated wool could considerably reduce the concentration of As in water and was estimated to have sufficient ability to trap a lethal dose of As. These findings show that the “affinity silk filter” is a potential device for the prophylaxis of aquatic animal diseases.
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