Comparison of whole-cell antigens of pressure- and formalin-killed Flexibacter columnaris from channel catfish (Ictalurus punctatus).
To identify and compare immunodominant antigens in whole-cell lysates of pressure- and formalin-killed Flexibacter columnaris. Sera from naturally infected and vaccinated channel catfish. Whole-cell lysates of pressure- and formalin-killed F columnaris were compared, and antigens were isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The antigens were identified by staining, western blotting, and specific monoclonal antibodies to glycoproteins. Western blotting was performed, using sera from channel catfish (Ictalurus punctatus) with naturally acquired F columnaris infection and sera from channel catfish vaccinated with an experimental prototype F columnaris vaccine. Whole-cell lysates of pressure and formalin-killed F columnaris shared 4 proteins: 100, 80, 66, and 60 kd. The 60-kd antigen was a glycoprotein. Western blotting, using sera from naturally infected channel catfish, revealed the same proteins for pressure- and formalin-killed F columnaris. Sera from vaccinated fish reacted only to pressure-killed lysate antigens. Pressure- and formalin-killed F columnaris whole-cell lysates share 100-, 80-, 66-, and 60-kd proteins and are recognized by antibodies from naturally infected catfish and those vaccinated with formalin-killed F columnaris. Formalin treatment modifies or inactivates the 60-kd protein antigens, rendering them unrecognizable to antibodies from channel catfish naturally infected with F columnaris, suggesting that formalin-killed F columnaris may not be suitable for use as a bacterin against columnaris disease.