An atlas of the germ ball-cercaria-schistosomulum transition in Schistosoma mansoni, using confocal microscopy and in situ hybridisation.

Schistosomes are complex platyhelminth parasites with a genome comprising ∼12,000 protein-coding genes, three distinct generations, and at least seven distinct phenotypes. We chart here cellular and gene expression changes associated with development of the cercaria, in the intramolluscan daughter sporocyst, and its transformation into the skin stage schistosomulum upon infection of the mammalian host. We describe the morphology of the early daughter sporocyst and the increasing complexity of cellular organisation in germ balls as they rapidly develop into cercariae. We show how individual myocytes differentiate and combine to create the complex musculature of the head capsule and body wall. In situ hybridisation reveals that some transcripts encoding the secretory proteins, released during skin penetration, are expressed in gland-cell precursors very early in germ ball development. However, those for the projected anti-inflammatory protein Sm16-stathmin are widely expressed in germ ball tissues, suggesting the protein has intracellular functions. Transcripts for smkk7 are expressed in six cells of the larval body, while the KK7 protein is present throughout the peripheral nerve net, including sensory nerve bulbs, providing a marker for the nerve net in adult worms. We also note that the cercaria-schistosomulum transformation is accompanied by tissue remodelling without growth.

Exploring the Fasciola hepatica tegument proteome.

The surface tegument of the liver fluke Fasciola hepatica is a syncytial cytoplasmic layer bounded externally by a plasma membrane and covered by a glycocalyx, which constitutes the interface between the parasite and its ruminant host. The tegument's interaction with the immune system during the fluke's protracted migration from the gut lumen through the peritoneal cavity and liver parenchyma to the lumen of the bile duct, plays a key role in the fluke's establishment or elimination. However, little is known about proteins of the tegument surface or its secretions. We applied techniques developed for the blood fluke, Schistosoma mansoni, to enrich a tegument surface membrane preparation and analyse its composition by tandem mass spectrometry using new transcript databases for F. hepatica. We increased the membrane and secretory pathway components of the final preparation to ∼30%, whilst eliminating contaminating proteases. We identified a series of proteins or transcripts shared with the schistosome tegument including annexins, a tetraspanin, carbonic anhydrase and an orthologue of a host protein (CD59) that inhibits complement fixation. Unique to F. hepatica, we also found proteins with lectin, cubulin and von Willebrand factor domains plus 10 proteins with leader sequences or transmembrane helices. Many of these surface proteins are potential vaccine candidates. We were hampered in collecting tegument secretions by the propensity of liver flukes, unlike blood flukes, to vomit their gut contents. We analysed both the 'vomitus' and a second supernatant released from haematin-depleted flukes. We identified many proteases, some novel, as well as a second protein with a von Willebrand factor domain. This study demonstrates that components of the tegumental surface of F. hepatica can be defined using proteomic approaches, but also indicates the need to prevent vomiting if tegument secretions are to be characterised.