Biomphalaria snails are instrumental in transmission of the human blood fluke Schistosoma mansoni. With the World Health Organization's goal to eliminate schistosomiasis as a global health problem by 2025, there is now renewed emphasis on snail control. Here, we characterize the genome of Biomphalaria glabrata, a lophotrochozoan protostome, and provide timely and important information on snail biology. We describe aspects of phero-perception, stress responses, immune function and regulation of gene expression that support the persistence of B. glabrata in the field and may define this species as a suitable snail host for S. mansoni. We identify several potential targets for developing novel control measures aimed at reducing snail-mediated transmission of schistosomiasis.

Biomphalaria glabrata snails that display either resistant or susceptible phenotypes to the parasitic trematode, Schistosoma mansoni provide an invaluable resource towards elucidating the molecular basis of the snail-host/schistosome relationship. Previously, we showed that induction of stress genes either after heat-shock or parasite infection was a major feature distinguishing juvenile susceptible snails from their resistant counterparts. In order to examine this apparent association between heat stress and snail susceptibility, we investigated the effect of temperature modulation in the resistant snail stock, BS-90. Here, we show that, incubated for up to 4 hrs at 32°C prior to infection, these resistant snails became susceptible to infection, i.e. shedding cercariae at 5 weeks post exposure (PE) while unstressed resistant snails, as expected, remained resistant. This suggests that susceptibility to infection by this resistant snail phenotype is temperature-sensitive (ts). Additionally, resistant snails treated with the Hsp 90 specific inhibitor, geldanamycin (GA) after heat stress, were no longer susceptible to infection, retaining their resistant phenotype. Consistently, susceptible snail phenotypes treated with 100 mM GA before parasite exposure also remained uninfected. These results provide direct evidence for the induction of stress genes (heat shock proteins; Hsp 70, Hsp 90 and the reverse transcriptase [RT] domain of the nimbus non-LTR retrotransposon) in B. glabrata susceptibility to S. mansoni infection and characterize the resistant BS-90 snails as a temperature-sensitive phenotype. This study of reversing snail susceptibility phenotypes to S. mansoni provides an opportunity to directly track molecular pathway(s) that underlie the B. glabrata snail's ability to either sustain or destroy the S. mansoni parasite. Biomphalaria glabrata snails that are either resistant or susceptible to the parasite, Schistosoma mansoni, have been an invaluable resource in studies aimed at understanding the molecular basis of the snail/schistosome interaction. Schistosomes cause the chronic debilitating disease schistosomiasis. Thus, it is hoped that dissecting pathways that underlie the snail/schistosome relationship might translate into alternative control strategies that will include blocking transmission of the parasite at the snail-stage of its development. Induction of stress genes is a feature distinguishing early exposed juvenile susceptible NMRI snails from resistant BS-90 snail stocks. To further analyze this apparent involvement of stress induction and snail susceptibility, here we applied heat stress to the resistant BS-90 snail, enhancing induction of stress genes (Hsp 70, Hsp 90 and RT) prior to infection. Results showed these resistant snails became susceptible, indicating resistance as being a temperature sensitive phenotype in these snails. Stressed resistant snails treated with the Hsp 90 specific inhibitor, geldanamycin, prior to exposure, were, however, shown to maintain their refractory phenotype. Interestingly, inhibitor treated susceptible snails also became non-susceptible. Collectively, these data point to stress induction as an important early step in the ability of S. mansoni to infect juvenile B. glabrata snails.

Biomphalaria glabrata snails play an integral role in the transmission of Schistosoma mansoni, the causative agent for human schistosomiasis in the Western hemisphere. For the past two decades, tremendous advances have been made in research aimed at elucidating the molecular basis of the snail/parasite interaction. The growing concern that there is no vaccine to prevent schistosomiasis and only one effective drug in existence provides the impetus to develop new control strategies based on eliminating schistosomes at the snail-stage of the life cycle. To elucidate why a given snail is not always compatible to each and every schistosome it encounters, B. glabrata that are either resistant or susceptible to a given strain of S. mansoni have been employed to track molecular mechanisms governing the snail/schistosome relationship. With such snails, genetic markers for resistance and susceptibility were identified. Additionally, differential gene expression studies have led to the identification of genes that underlie these phenotypes. Lately, the role of schistosomes in mediating non-random relocation of gene loci has been identified for the first time, making B. glabrata a model organism where chromatin regulation by changes in nuclear architechture, known as spatial epigenetics, orchestrated by a major human parasite can now be investigated. This review will highlight the progress that has been made in using molecular approaches to describe snail/schistosome compatibility issues. Uncovering the signaling networks triggered by schistosomes that provide the impulse to turn genes on and off in the snail host, thereby controlling the outcome of infection, could also yield new insights into anti-parasite mechanism(s) that operate in the human host as well.

An in vivo, non-invasive technique for gene silencing by RNA interference (RNAi) in the snail, Biomphalaria glabrata, has been developed using cationic polymer polyethyleneimine (PEI) mediated delivery of long double-stranded (ds) and small interfering (si) RNA. Cellular delivery was evaluated and optimized by using a ‘mock’ fluorescent siRNA. Subsequently, we used the method to suppress expression of Cathepsin B (CathB) with either the corresponding siRNA or dsRNA of this transcript. In addition, the knockdown of peroxiredoxin (Prx) at both RNA and protein levels was achieved with the PEI-mediated soaking method. B. glabrata is an important snail host for the transmission of the parasitic digenean platyhelminth, Schistosoma mansoni that causes schistosomiasis in the neotropics. Progress is being made to realize the genome sequence of the snail and to uncover gene expression profiles and cellular pathways that enable the snail to either prevent or sustain an infection. Using PEI complexes, a convenient soaking method has been developed, enabling functional gene knockdown studies with either dsRNA or siRNA. The protocol developed offers a first whole organism method for host-parasite gene function studies needed to identify key mechanisms required for parasite development in the snail host, which ultimately are needed as points for disrupting this parasite mediated disease. Freshwater snails are important in the transmission of schistosomiasis. As part of an integral control effort to combat the spread of schistosomiasis new intervention tools are being sought. One method is to interrupt the transmission of the causative schistosome parasite during the intra-molluscan phase of its development. Gene-silencing technology involving the use of dsRNA have used an injection route to disrupt gene translation in the Schistosoma mansoni snail host, Biomphalaria glabrata in an effort to investigate how inhibition of various transcripts can affect the dynamics of the snail/parasite interaction. These studies have been helpful in showing us that a gene-silencing pathway that uses dsRNA indeed exists in snails but the injection method previously utilized is impractical, especially when working with juvenile snails. To make the use of gene silencing technology more widely applicable to functional gene studies in snails, we have developed a more convenient soaking method that uses a cationic carrier polyethylene amine (PEI) to deliver dsRNA or siRNA into juvenile snails. Using this method we show the successful knockdown at both RNA and protein levels of the B. glabrata peroxiredoxin (Prx) gene. The method was also evaluated for silencing the Cathepsin B (CathB) gene in the snail.