Schistosoma mansoni: quantification of skin penetration and early migration by differential external radioassay and autoradiography

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Abstract
SUMMARY: Eleven gamma-emitting radionuclides (49Sc, 54Mn, 59Fe ( + 2 and + 3), 60Co, 65Zn, 75Se (as selenomethionine, selenocystine, selenite and selenate), 109Cd, 125Sb, 133Ba, 137Cs and 203Hg) were screened as labelling agents for Schistosoma mansoni cercariae by incubation of infected Biomphalaria glabrata snails in radioactive solution according to the technique of Christensen (1977). Only [75Se]methionine yielded satisfactorily labelled cercariae. Multiple regression analysis of volume, number of cercariae and radioactivity from a series of 10 aliquots of unwashed cercarial suspensions yielded estimates of unbound and cercarial-bound radioactivity that were equivalent or superior to estimates based on assay of washed cercariae and eliminated loss of cercariae. Washing of cercarial suspensions over 8 μm pore diameter Millipore filters was found to result in entanglement of 60–90% of the cercariae on the filter disc. Differential external radioassay, a new technique employing partial body shielding within a total body counter, permitted separate estimation of tail and body radioactivity of conscious mice previously exposed by tail immersion to 75Se-labelled cercariae, with measurements repeated as often as desired. Approximately 39% of the 75Se present in emergent cercariae was retained by schistosomula transformed in vitro but this was subject to considerable variation, especially in schistosomula transformed in vivo. Secreted or catabolized label from penetrant cercariae and schistosomula was rapidly removed from the skin by the bloodstream. Numbers of schistosomula in tail skin were directly proportional to the number of reduced silver foci counted on tail autoradiograms; only a very small fraction of tail radioactivity represented unbound (‘spurious’) label. Migration of schistosomula away from skin was 50% complete at 3·8–4·3 days, as determined by probit analysis of autoradiographic data.