Tegument surface membranes of adult Schistosoma mansoni: development of a method for their isolation

Abstract

Several approaches to surface membrane stripping have been applied to the adult schistosome. Membrane removal was evaluated by the use of different extrinsic and intrinsic markers of which alkaline phosphatase proved to be the most reliable. After initial studies employing incubation of worms in buffer alone, Triton X-100 or freeze/thaw, the last method was chosen for development. The final method applies a single freeze/thaw step to adult worms in balanced salt solution followed by short bursts of agitation on a vortex mixer to release the tegument. Differential and density gradient steps subsequently yield a final membrane pellet enriched over 130 times in surface alkaline phosphatase. The method has been characterized during its development using electron microscopy and enzyme markers for contaminant worm fractions.