Molecular techniques can aid in the classification of Biomphalaria species because morphological differentiation between these species is difficult. Previous studies using phylogeny, morphological and molecular taxonomy showed that some populations studied were Biomphalaria cousini instead of Biomphalaria amazonica. Three different molecular profiles were observed that enabled the separation of B. amazonica from B. cousini. The third profile showed an association between the two and suggested the possibility of hybrids between them. Therefore, the aim of this work was to investigate the hybridism between B. cousini and B. amazonica and to verify if the hybrids are susceptible to Schistosoma mansoni. Crosses using the albinism factor as a genetic marker were performed, with pigmented B. cousini and albino B. amazonica snails identified by polymerase chain reaction-restriction fragment length polymorphism. This procedure was conducted using B. cousini and B. amazonica of the type locality accordingly to Paraense, 1966. In addition, susceptibility studies were performed using snails obtained from the crosses (hybrids) and three S. mansoni strains (LE, SJ, AL). The crosses between B. amazonica and B. cousini confirmed the occurrence of hybrids. Moreover, hybrids can be considered potential hosts of S. mansoni because they are susceptible to LE, SJ and AL strains (4.4%, 5.6% and 2.2%, respectively). These results indicate that there is a risk of introducing schistosomiasis mansoni into new areas.

The present study was aimed at characterising Biomphalaria species using both morphological and molecular (PCR-RFLP) approaches. The specimens were collected in 15 localities in 12 municipalities of the southern region of the state of Rio Grande do Sul, Brazil. The following species were found and identified: Biomphalaria tenagophila guaibensis, Biomphalaria oligoza and Biomphalaria peregrina. Specimens of the latter species were experimentally challenged with the LE Schistosoma mansoni strain, which showed to be refractory to infection.

Resistant (Taim, RS) and susceptible albino (Joinville, SC) Biomphalaria tenagophila populations were kept together, at different proportions, throughout a 18-month-period. Some of the snail groups were submitted to Schistosoma mansoni infection. The targets of this study were (a) to analyze the populational dynamics among resistant and susceptible individuals to S. mansoni; (b) to study the resistance phenotype in descendants of cross-breeding; (c) to observe whether the parasite could exert any kind of selection in those snail populations. Throughout the experiment it could be observed that the susceptible B. tenagophila strain (Joinville) underwent a selective pressure of the parasite that was negative, since the individuals showed a high mortality rate. Although B. tenagophila (Taim) population presented a higher mortality rate without pressure of the parasite, this event was compensated by a reproductive capacity. B. tenagophila Taim was more fecund than B. tenagophila Joinville and was able to transmit the resistance character to their descendants. F1 generation obtained by cross-breeding between resistant and susceptible lineages was completely resistant to S. mansoni infection, irrespective of the Taim proportion. Moreover, less than 5% of F2 progeny were susceptible to S. mansoni infection.

This is the first report on occurrence of Biomphalaria straminea in the district of São José de Almeida (municipality of Jaboticatubas) State of Minas Gerais, Brazil. The presence of B. glabrata and B. tenagophila had already been reported in this area. Such municipality is part of the metropolitan region of Belo Horizonte and comprises 60% of the Tourist Complex of Serra do Cipó. Since the 1950s throughout the 1990s, a schistosomiasis prevalence ranging from 15 to 40% has been observed. Although no B. straminea specimen has been found naturally infected in the region, descendants of these snails collected in the area, showed to be experimentally susceptible to Schistosoma mansoni infection reaching rates from 14.6 to 28.6%. Even not being found naturally infected, in the State of Minas Gerais, the possibility that the species B. straminea may keep endemicity foci of schistosomiasis should be regarded, as in the Northeastern region of Brazil where the high density of this planorbid and the social-economic and sanitary conditions enable to the transmission.

Para determinar a taxa de infecção pelo Schistosoma mansoni em Biomphalaria tenagophila e em B. straminea foi utilizada a reação em cadeia da polimerase em baixa estringência (LS-PCR), como técnica complementar ao método de exposição à luz. A LS-PCR já foi padronizada no nosso laboratório para detectar o DNA do trematódeo em B. glabrata. A taxa de infecção pelo S. mansoni foi maior quando utilizou-se a técnica convencional e a LS-PCR. O perfil do DNA do parasita foi detectado após 7 dias de exposição a miracídios em ambas as espécies, quando utilizou-se a LS-PCR. Esta técnica possibilita a detecção precoce de focos de transmissão, em áreas endêmicas, antes do início da eliminação de cercárias.

In order to determine Schistosoma mansoni infection rates in Biomphalaria tenagophila and B. straminea, low stringency polymerase chain reaction (LS-PCR) technique was used as a complementary method to light exposure technique. LS-PCR has already been standardized in our laboratory to detect the trematode DNA in B. glabrata. Higher S. mansoni infection rates were detected using conventional method and LS-PCR. The parasite DNA profile was detected in both species after 7-day exposure to miracidia, using LS-PCR. This technique enables early detection of schistosomiasis transmission focuses, in endemic areas, before the beginning of cercariae shedding.

Um levantamento malacológico em coleções hídricas de 13 municípios da microrregião de Belo Horizonte, MG, Brasil, foi efetuado para detectar focos de transmissão de esquistossomose e outras parasitoses. De 1990 a 1996 foram coletados 22.066 moluscos dos quais 378 (1,7%) estavam infectados com trematódeos: Biomphalaria glabrata (7.920), com Schistosoma mansoni (1,9%), com Echinostomatidae (1,2%), com Strigeidae (0,6%), com Cercaria minense (0,1%) e Derogenidae (-0,1%); B. straminea (4.093), com Strigeidae (0,6%), com Echinostomatidae (0,2%), com Clinostomatidae (-0,1%) e duas cercárias desconhecidas; B. tenagophila (1.338), com Strigeidae (0,1%); Physa marmorata (1.776), com Echinostomatidae (1,6%). Os moluscos Biomphalaria peregrina, B. occidentalis, B. schrammi, Drepanotrema depressissimum, D. lucidum, D. cimex, Physa cubensis, Lymnaea columella, Melania tuberculata, Idiopyrgus souleyetianus, Pomacea sp, Anodontites sp e Ancylidae não estavam infectados. Moluscos de 9 municípios estavam infectados com S. mansoni e de 11 com outros trematódeos.

A malacological survey to detect foci of transmission of schistosomiasis and other parasitic diseases was undertaken into water-courses from 13 municipalities of microregion of Belo Horizonte, MG, Brazil. From 1990 to 1996, 22,066 snails were collected. From those, 378 (1.7%) were found infected by trematodes: Biomphalaria glabrata (7,920), infected by Schistosoma mansoni (1.9%), Echinostomatidae (1.2%), Strigeidae (0.6%), Cercaria minense (0.1%) and Derogenidae (-0.1%); B. straminea (4,093) infected by Strigeidae (0.6%), Echinostomatidae (0.2%), Clinostomatidae (-0.1%) and two unidentified cercariae; B. tenagophila (1,338), infected by Strigeidae (0.1%) and Physa marmorata (1,776) by Echinostomatidae (1.6%). The snails Biomphalaria peregrina, B. occidentalis, B. schrammi, Drepanotrema depressissimum, D. lucidum, D. cimex, Physa cubensis, Lymnaea columella, Melania tuberculata, Idiopyrgus souleyetianus, Pomacea sp, Anodontites sp and Ancylidae were found noninfected. Snails from 9 municipalities were infected by S. mansoni and from 11 by other trematodes.

A densidade planorbídica, as taxas de infecção e a produção mensal de cercárias de Schistosoma mansoni por Biomphalaria glabrata, foram determinadas em foco do Barreiro de Baixo (Belo Horizonte MG, Brasil). Durante 38 meses, de 1984 a 1987, foram capturados 5.366 moluscos dentre os quais 324 (6,03%) estavam infectados com o S. mansoni. O total de cercárias eliminadas foi de 5.667.312. Cada molusco eliminou em média 17.422 cercárias durante sua permanência no laboratório. A maior longevidade dos moluscos infectados foi de 218 dias. Foi observada a cura natural de 42 (12,9%) dos exemplares infectados, após cerca de 130 dias de captura. A densidade planorbídica média, no foco durante esse período foi de 16,3 moluscos por conchada. A eliminação de cercárias pelos moluscos do campo foi comparada com a de exemplares criados no laboratório, infectados em massa com a linhagem LE de S. mansoni de Belo Horizonte. Os moluscos do laboratório eliminaram em média 6.061 cercárias por exemplar, número 2,8 vezes menor do que os moluscos do campo, devido a longevidade menor. A taxa de prevalência da esquistossomose no foco foi de 14,3%.

The snail density, levels of infection and the monthly production of Schistosoma mansoni cercariae by Biomphalaria glabrata were determined in a focus of Barreiro de Baixo (Belo Horizonte, MG, Brazil). During a period of 38 months (1984 to 1987) 5,366 snails were collected of which 324 (6.03%) were infected with S. mansoni. The total number of cercariae shed was 5,667,312. Each snail shed an average of 17,422 cercariae during the time that it was under study in the laboratory. The greatest longevity of infected snails was 218 days. Natural cure was observed in 42 (12.9%) of the infected specimens about 130 days after collection. The average snail density in the focus during the period of study was 16.3 snails per scoop. The shedding of cercariae by snails collected from the field was compared with laboratory bred specimens infected in mass with the LE strain of S. mansoni from Belo Horizonte. The laboratory infected snails shed an average of 6,061 cercariae each, a value 2.8 times less than the field specimens due to a shorter life span. The prevalence of schistosomiasis in the focus was 14.3%.

Bioensaios usando extratos hexânicos de cascas da castanha do caju, Anacardium occidentale, coletadas no Ceará em 1972 (amostra 1) e em 1987 (amostra 2) foram feitos sobre moluscos adultos e desovas de Biomphalaria glabrata, B. tenagophila e B. straminea, no laboratório e no campo. As cascas, 18,5 g, sem triturar amostra 1, também foram testadas sobre moluscos adultos e desovas das três espécies. A toxidez do extrato foi testada ainda sobre peixes (Poecilia reticulata) e girinos. As concentrações letais CL90, amostra 1, foram de 2,0 a 2,2 ppm para os moluscos das três espécies. Para B. glabrata adultos, recém-eclodidos e desovas as CL90, amostra 2, foram de 2,0, 0,5 e 30,0 ppm respectivamente. As cascas causaram mortalidade de 40 a 80% dos moluscos e de 22 a 35% dos embriões, ocasionando redução de 40 a 55% na oviposição das três espécies. O extrato hexânico, amostra 2, foi inócuo para girinos e peixes até 2 ppm. No campo, em poços com água parada, tratados com 20 ppm do extrato, amostra 1, ocorreu 97,1% de mortalidade de B. straminea e 100% de B. glabrata e B. tenagophila. Com a niclosamida a 3 ppm ocorreu 100% de mortalidade das três espécies.

Bioassays using hexanolic extracts of cashew nut shells, of Anacardium occidentale, collected in Ceará in 1972 (Sample 1) and 1987 (Sample 2) were undertaken with adult snails and egg masses of Biomphalaria glabrata, B. tenagophila and B. straminea both in the laboratory and in the field. Non extracted shells, 18.5 g, sample 1, were also tested with adult snails and egg masses of the three species. The toxicity of extract was tested with fish (Poecilia reticulata) and tadpoles. The lethal concentration, CL90, of sample 1 was from 2.0 to 2.2 ppm for adult snails of the three species. With sample 2, the CL90 was 2.0, 0.5 and 30.0 ppm for B. glabrata adults, newly hatched snails and egg mass respectively. Non extracted shells caused 40 - 80% mortality of adult snails, 22 - 35% mortality of embryos and 40 - 55% reduction of egg production. The hexanolic extract, sample 2, were inocuous for tadpole and fish at 2 ppm. In the field, in pools of still water treatment with 20 ppm of extract, sample 1, caused a 97.1% mortality of B. straminea and 100% mortality of B. glabrata and B.tenagophila. Using Niclosamide, at 3 ppm, 100% mortality of the three species occured.