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Diseases of Aquatic Organisms

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DAO 106:149-162 (2013)  -  DOI:

Antigenic characterization of Enteromyxum leei (Myxozoa: Myxosporea)

Itziar Estensoro*, Pilar Álvarez-Pellitero, Ariadna Sitjà-Bobadilla

Instituto de Acuicultura Torre de la Sal, Consejo Superior de Investigaciones Científicas (IATS-CSIC), Torre de la Sal s/n, 12595 Ribera de Cabanes, Castellón, Spain

ABSTRACT: Enteromyxum leei, an intestinal myxozoan parasite affecting a wide range of fish, was partially purified, and the immunogenic composition of its glycoproteins as well as the proteolytic activity were studied. Parasite extracts, mainly containing spores, were separated by SDS-PAGE, and thereafter, immunoblots were carried out with a polyclonal antiserum (Pab) raised against E. leei. Periodic acid/Schiff staining of gels, periodate- and Proteinase K-treated Western blots and Lectin blots were performed to analyse the terminal carbohydrate composition of the parasite’s antigens. Additionally, the cross-reaction of the parasite extracts with a Pab raised against the polar filament of the myxozoan Myxobolus pendula was studied. Both Pabs detected proteic epitopes on antigenic proteins and glycoproteins of E. leei, ranging between 15 and 280 kDa. In particular, 2 glycoproteic bands (15 and 165 kDa), immunoreactive to both Pabs and with glucose and mannose moieties, could correspond to common antigens shared among myxozoans. The 165 kDa band also presented galactose, N-acetyl-galactosamine and N-acetyl-glucosamine, pointing to its possible origin on chitin-built spore valves and to its possible involvement in host-parasite interactions. The molecular weight of the 15 kDa glycoproteic antigen matches that of minicollagen, a cnidarian-specific protein of nematocysts with a myxozoan homologue. Several proteases with apparent molecular weights ranging between 43 and 245 kDa were found in zymographies of E. leei extracts, and these may have a potential role in the parasite’s pathogenesis. This is a useful approach for further studies to detect targets for antiparasitic therapy.

KEY WORDS: Western blot · Polyclonal antibody · Glycoprotein · Lectin · Protease · Parasite

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Cite this article as: Estensoro I, Álvarez-Pellitero P, Sitjà-Bobadilla A (2013) Antigenic characterization of Enteromyxum leei (Myxozoa: Myxosporea). Dis Aquat Org 106:149-162.

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