Genetic variability and identification of the intermediate snail hosts of Schistosoma mansoni

Studies based on shell or reproductive organ morphology and genetic considerations suggest extensive intraspecific variation in Biomphalaria snails. The high variability at the morphological and genetic levels, as well as the small size of some specimens and similarities between species complicate the correct identification of these snails. Here we review our work using methods based on polymerase chain reaction (PCR) amplification for analysis of genetic variation and identification of Biomphalaria snails from Brazil, Argentina, Uruguay and Paraguay. Arbitrarily primed-PCR revealed that the genome of B. glabrata exihibits a remarkable degree of intraespecific polymorphism. Low stringency-PCR using primers for 18S rRNA permited the identification of B. glabrata, B. tenagophila and B. occidentalis. The study of individuals obtained from geographically distinct populations exhibits significant intraspecific DNA polymorphism, however specimens from the same species, exhibit some species specific LSPs. We also showed that PCR-restriction fragment of length polymorphism of the internal transcribed spacer region of Biomphalaria rDNA, using DdeI permits the differentiation of the three intermediate hosts of Schistosoma mansoni. The molecular biological techniques used in our studies are very useful for the generation of new knowledge concerning the systematics and population genetics of Biomphalaria snails.

Molecular characterisation of intermediate snail hosts and the search for resistance genes

The relationship between schistosomes and their intermediate hosts is an extremely intricate one with strains and species of the parasite depending on particular species of snail, which in turn may vary in their susceptibility to the parasites. In order to gain a better understanding of the epidemiology of the disease we have been investigating the use of molecular markers for snail identification and for studying host-parasite relationships. In this paper we will draw on examples concerning schistosomiasis in West and East Africa to illustrate how a molecular analysis can be used as part of a "total evidence" approach to characterisation of Bulinus species and provide insights into parasite transmission. Particular emphasis is given to ribosomal RNA genes (rRNA), random amplified polymorphic DNA (RAPDs) and the mitochondrial gene cytochrome oxidase I (COI). Snails resistant to infection occur naturally and there is a genetic basis for this resistance. In Biomphalaria glabrata resistance to Schistosoma mansoni is known to be a polygenic trait and we have initiated a preliminary search for snail genomic regions linked to, or involved in, resistance by using a RAPD based approach in conjunction with progeny pooling methods. We are currently characterising a variety of STSs (sequence tagged sites) associated with resistance. These can be used for local linkage and interval mapping to define genomic regions associated with the resistance trait. The development of such markers into simple dot-blot or specific PCR-based assays may have a direct and practical application for the identification of resistant snails in natural populations.

Possible hybridization of Brazilian planorbid snails and its importance in population dynamics

This study focuses on the possibility of experimental hybridization among host snail species for Schistosoma mansoni in Brazil, with morphological characterization of the hybrids found. By using albinism as a genetic marker, intraspecific crossbreedings were performed between two strains of each species involved, in addition to interspecific crossbreedings; the only viable crossbreeding was between pigmented Biomphalaria glabrata (Paulista, PE) and albino B. tenagophila (Joinville, SC), with the formation of F1 and F2 generations. All offspring in F1 displayed black eyes and a renal ridge on the mantle, while F2 displayed dissociated morphological traits. With regard to reproduction, F1 was more efficient than F2. The experiment's results suggest post-zygotic reproductive isolation.

Susceptibility of Nectomys rattus (Pelzen, 1883) to experimental infection with Schistosoma mansoni (Sambon, 1907): a potential reservoir in Brazil

The aim of the present research was to evaluate the potential of Nectomys rattus, the "water rat", to develop Schistosoma mansoni infection. Comparison with N. squamipes was carried out. Both species of rodents were submitted to transcutaneous infection using different infective cercariae loads: 50, 100 or 500. N. rattus showed high susceptibility to S. mansoni, with an infection rate of 71%. Rodents were able to excrete viable eggs of S. mansoni in the feaces during all infection period. For both species, the small intestine, followed by the liver and the large intestine, presented the highest concentration of eggs among the surveyed organs. Infection caused no animal death. Moreover, N. rattus accomplished the parasite's life cycle, by infecting the snails Biomphalaria glabrata and later Mus musculus. These evidences indicate that both N. rattus, as for N. squamipes are potential reservoirs for schistosomiasis in Brazil. Considering the fact that N. rattus and N. squamipes exist in the same natural ecosystems of S. mansoni, we suggest that these rodents must be regarded as influential factors in epidemiology surveys.

The Last Fifteen Years of Schistosomiasis in Venezuela: Features and Evolution

Control of schistosomiasis in Venezuela has been a topic of major interest and controversy among the metaxenic parasitosis. A small area of transmission of approximately 15,000 km2 was thought to be eradicated some years ago. However, some epidemiological characteristics of our transmission area have limited the success on the way toward eradication. Since 1945, when the Schistosomiasis Control Program started, the prevalence in the endemic area has decreased from 14% in 1943 to 1.4% in 1996. Until 1982, the surveillance of active cases was based on massive stool examination. Since then, the Schistosomiasis Research Group (SRG) recommended the additional use of serologic tests in the Control Program and the selective or massive chemotherapy depending on serological and parasitological prevalence of each community. At present, the real prevalence is underestimated due to the fact that approximately 80% of the individuals eliminate less than 100 eggs/g of feces. Those persons could be responsible for the maintenance of the foci going on and therefore limiting the impact of the control measures. Efforts of the SRG are being oriented toward improvement of immunodiagnostic tests by using defined antigens (enzymes) and chemically synthesized peptides, derived from relevant molecules of the parasite, either for antibodies or antigens search. On the other hand, introduction of snail competitors has been a biological weapon in the control of the intermediate host in certain areas. However, the recent reinfestation of water courses by Biomphalaria glabrata, the increased prevalence in some areas, together with important administrative changes at the Control Program of the Minister of Health, have arisen new questions and doubts, challenging the eradication strategy proposed during the last decade.

Identification of planorbids from Venezuela by polymerase chain reaction amplification and restriction fragment length polymorphism of internal transcriber spacer of the RNA ribosomal gene

Snails of the genus Biomphalaria from Venezuela were subjected to morphological assessment as well as polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) analysis. Morphological identification was carried out by comparison of characters of the shell and the male and female reproductive apparatus. The PCR-RFLP involved amplification of the internal spacer region ITS1 and ITS2 of the RNA ribosomal gene and subsequent digestion of this fragment by the restriction enzymes DdeI, MnlI, HaeIII and MspI. The planorbids were compared with snails of the same species and others reported from Venezuela and present in Brazil, Cuba and Mexico. All the enzymes showed a specific profile for each species, that of DdeI being the clearest. The snails were identified as B. glabrata, B. prona and B. kuhniana.

Biological screening of Brazilian medicinal plants

In this study, we screened sixty medicinal plant species from the Brazilian savanna ("cerrado") that could contain useful compounds for the control of tropical diseases. The plant selection was based on existing ethnobotanic information and interviews with local healers. Plant extracts were screened for: (a) molluscicidal activity against Biomphalaria glabrata, (b) toxicity to brine shrimp (Artemia salina L.), (c) antifungal activity in the bioautographic assay with Cladosporium sphaerospermum and (d) antibacterial activity in the agar diffusion assay against Staphylococcus aureus, Escherichia coli, Bacillus cereus and Pseudomonas aeruginosa. Forty-two species afforded extracts that showed some degree of activity in one or more of these bioassays.

Invading freshwater snails and biological control in Martinique Island, French West Indies

Eight alien freshwater snail species were introduced into Martinique Island during the last 50 years. The introduced snails include four planorbids (Biomphalaria straminea, Helisoma duryi, Amerianna carinata and Gyraulus sp.), three thiarids (Melanoides tuberculata, M. amabilis and Tarebia granifera) and one ampullarid (Marisa cornuarietis). Four of these species rapidly colonized the whole Martinican hydrographic system whereas the other four remained restricted to some particular sites. The invasion processes were documented during the last 20 years and showed (i) a rapid invasion of the island by several morphs of M. tuberculata at the beginning of the 80's; (ii) the introduction of T. granifera in 1991 and M. amabilis in 1997; and (iii) the rapid spread of these last two species throughout the island. In the years following its introduction, M. tuberculata was used in biological control experiments against the snail hosts of schistosomiasis, B. glabrata and B. straminea. Experiments were conducted with success in several groups of water-cress beds which constituted the latest transmission sites for schistosomiasis at the beginning of the 80's. A malacological survey carried out in 2000 all over the island showed the absence of B. glabrata but the presence of some residual populations of B. straminea. Long-term studies carried out in Martinique have shown that the thiarids are able to maintain relatively stable populations over a long period of time, thus preventing recolonization by the snail hosts. Within this context the invasion of the hydrographic system of Martinique by thiarid snails has resulted in an efficient and sustainable control of the intermediate hosts of schistosomiasis.

Studies on the biology of schistosomiasis with emphasis on the Senegal river basin

The construction of the Diama dam on the Senegal river, the Manantali dam on the Bafing river, Mali and the ensuing ecological changes have led to a massive outbreak of Schistosoma mansoni in Northern Senegal, associated with high intensity of infections, due to intense transmission, and the creation of new foci of S. haematobium. Data on the vectorial capacity of Biomphalaria pfeifferi from Ndombo, near Richard Toll, Senegal are presented with sympatric and allopatric (Cameroon) S. mansoni. Comparisons are made on infectivity, cercarial production, chronobiology of cercarial emergence and longevity of infected snails. Recent data on the intermediate host specificity of different isolates of S. haematobium from the Lower and Middle Valley of the Senegal river basin (SRB) demonstrate the existence of at least two strains of S. haematobium. The role of Bulinus truncatus in the transmission of S. haematobium in the Lower and Middle Valleys of the SRB is reviewed. Both S. haematobium and S. mansoni are transmitted in the same foci in some areas of the SRB.

Possible competitive displacement of planorbids by Melanoides tuberculata in Minas Gerais, Brazil

Several species of snails, including Pomacea haustrum, Marisa cornuarietis and Helisoma duryi, have been identified as probable competitors and/or predators of planorbid intermediate hosts of Schistosoma. During the last few years, studies carried out in the Caribbean region have shown reductions and even disappearances of populations of Biomphalaria glabrata and B. straminea in breeding places where the snail Melanoides tuberculata was introduced. Observations made over a period of 10 years in two lakes close to Belo Horizonte, MG, showed that there were marked reductions in autochthonous populations of B. glabrata and B. straminea after the arrival of M. tuberculata, both Biomphalaria species disappearing completely after eight years.

Freshwater snails and schistosomiasis mansoni in the State of Rio de Janeiro, Brazil: I - metropolitan mesoregion

In order to elaborate a planorbid chart of the State of Rio de Janeiro a survey of freshwater gastropods in the Metropolitan Mesoregion of this State was performed and revealed the occurrence of 20 species: Antillorbis nordestensis (Lucena, 1954); Biomphalaria glabrata (Say, 1818); Biomphalaria schrammi (Crosse, 1864); Biomphalaria straminea (Dunker, 1848); Biomphalaria tenagophila (Orbigny, 1835); Burnupia sp.; Drepanotrema anatinum (Orbigny, 1835); Drepanotrema cimex (Moricand, 1839); Drepanotrema lucidum (Pfeiffer, 1839); Ferrissia sp.; Gundlachia ticaga (Marcus & Marcus, 1962); Heleobia davisi Silva & Thomé, 1985; Lymnaea columella Say, 1817; Melanoides tuberculatus (Müller, 1774); Physa cubensis Pfeiffer, 1839; Physa marmorata Guilding, 1828; Pomacea sp.; Pomacea canaliculata (Lamarck, 1822); Pomacea lineata (Spix, 1827) and Pomacea sordida (Swainson, 1823). Among the planorbid species B. tenagophila was the most frequent, occurring in all municipalities surveyed. The present study extends the distribution of B. straminea in the State of Rio de Janeiro and reports new records for A. nordestensis, B. schrammi, G. ticaga, H. davisi and the genera Burnupia and Ferrissia. An account about the current transmission areas of schistosomiasis mansoni in this Mesoregion is presented as well.

Freshwater snails and Schistosomiasis mansoni in the State of Rio de Janeiro, Brazil: II - Centro Fluminense Mesoregion

During the course of a survey carried out from 2000 to 2001 in the Centro Fluminense Mesoregion of the State of Rio de Janeiro 22 molluscan species were found. Many of the records are new due to the dearth of previous studies. Concerning the snail hosts of Schistosoma mansoni, the most frequently encountered species was Biomphalaria tenagophila, as it occurred in all the surveyed municipalities. There are new records of Biomphalaria straminea and Biomphalaria peregrina which is regarded as a potential intermediate host. Drepanotrema lucidum and Antillorbis nordestensis were found to be shedding echinostome cercariae and strigid cercariae respectively. An account about the current schistosomiasis transmission sites in this Mesoregion is presented as well.