Time series analysis of tegument ultrastructure of in vitro transformed miracidium to mother sporocyst of the human parasite Schistosoma mansoni

Abstract

The transformation of Schistosoma mansoni miracidia into mother sporocysts is induced, either in vivo by the penetration of the free-living larval stage, the miracidium, in the snail Biomphalaria glabrata or in vitro following the incubation of the miracidium in Chernin's Balanced Salt Solution (CBSS) or Bge (B. glabrata embryonic cell line) culture medium. The in vitro development of S. mansoni miracidium into mother sporocyst was monitored by Scanning Electron Microscopy (SEM) from 2.5 h to 120 h in CBSS. The transformation starts when the miracidium ciliate plates detach due to the proliferation of the intercellular ridge associated with the degeneration of mid-body papillae of the miracidium. The loss of ciliated plates causes the appearing of scars, filled across time by the proliferation of a new tegument originating from the interplate ridge. This new tegument covers the entire body of the metamorphosing parasite and differentiates over time, allowing some exchanges (uptakes or secretion/excretion) between the parasite and its host. In contrast to the well-described development of adult and free-living larval stages of S. mansoni using SEM, the developmental transformation of intramolluscan stages, especially tegumental changes in the mother sporocyst, has been sparcely documented at the ultrastructural level. In addition, taking into account the latest literature on miracidium electron microscopy and the advances in SEM technologies over the last thirty years, the present study gathers three main objectives: (i) Fill the gap of tegument scanning electron micrographs of in vitro transforming sporocysts; (ii) Update the current bibliographic miracidia and sporocysts image bank due to rapid evolution of SEM technology; (iii) Understand and describe the critical steps and duration of the in vitro miracidium-to-sporocyst transformation process to assist in understanding the interaction between the larval surface and snail immune factors.