DAO 40:117-124 (2000)  -  doi:10.3354/dao040117

ABSTRACT: The biochemical composition of Sphaerospora dicentrarchi was studied. Periodate and Proteinase K treatments as well as lectin blots were used to analyse carbohydrate terminals. Zymography was applied to detect proteases. Four polyclonal antisera, raised against S. dicentrarchi (RaSdic), S. testicularis (RaStest), Ceratomyxa labracis (RaClab) and C. sparusaurati (RaCspr), were used in SDS polyacrylamide gel electrophoresis and immunoblot. Bands with molecular weight (MW) between 32 and 130 kDa were detected by electrophoresis. After Proteinase K treatment, apparent digestion of bands heavier than 43 kDa took place. RaSdic and RaStest detected similar bands with MW between 20 and 50 kDa, whereas RaClab and RaCspr recognized bands between 50 and 140 kDa. The 50 kDa band was recognized by all the polyclonal antisera, suggesting that it could correspond to an antigen shared by several myxosporean parasites. Four proteases were observed by zymography. From the 5 lectins assayed, binding was only observed using Con-A, which detected 2 bands of 96 and 78 kDa. Periodate treatment did not produce any effect on the binding of RaSdic and RaStest, but a high decrease of intensity in the antibody binding occurred at a concentration of 10 and 20 mM periodate when RaClab and RaCspr were tested. These results give information on the antigenic composition of S. dicentrarchi which could be useful for further diagnostic or immunoprevention studies.

KEY WORDS: Myxozoa · Sphaerospora · Ceratomyxa · Immnunoblotting · Glycoconjugates · Lectins · Zymography