Biomphalaria occidentalis Paraense, 1981 from Varzea das Flores dam, MG, Brazil, was exposed to infection with Schistosoma mansoni. Individual infection was performed with 140 B. occidentalis and 100 B. glabrata snails using LE and SJ strains. Two groups of B. occidentalis were killed after seven day-miracidia exposure to detect S. mansoni DNA, through the low stringency polymerase chain reaction (LS-PCR), and were negative. The infection rates were 69.2% (LE strain) and 96.7% (SJ strain) for B. glabrata and 0% for B. occidentalis. LS-PCR enabled early resistance diagnosis.

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.

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.

A comparative histopathological study of three snails species - Biomphalaria glabrata, B. tenagophila and B. straminea - which had been infected with Schistosoma mansoni miracidia revealed similar qualitative features, consisting of areas of sporocyst proliferation and differentiation associated with reactive host reaction, at the time they were actively eliminating great number of cercariae. However, in specimens that were exposed to miracidia but failed to eliminate cercariae later on, different histopathological pictures were observed in different snail species. While B. glabrata exhibited frequent focal (granulomatous) proliferation of amebocytes in several organs, B. tenagophila and B. straminea only rarely showed such reactive changes, suggesting that the mechanism of resistance to miracidial infection probably follows different pathways in the snail species studied

Foi feito estudo comparativo do desenvolvimento do Schistosoma mansoni na fase intra-molusco, através de cortes histológicos, em Biomphalaria tenagophila, B. straminea e B. glabrata. Duzentos moluscos de cada espécie foram expostos individualmente a 50 miracídios de S. mansoni da linhagem AL. Nenhuma larva foi observada nos exemplares fixados 72 horas após a exposição. Nos exemplares eliminando cercárías, 31 dias após a exposição, foram observadas reações teciduais de encapsulamento de larvas em B. tenagophila e B. straminea, na região cefalopodal, colar do manto e dutos renais. Nas glândulas digestivas das duas espécies não foram observadas reações. Em B. glabrata foi registrada a presença de numerosos esporocistos e cercárias sem reação tecidual na glândula digestiva e outros órgãos. As taxas de infecção desses moluscos com AL e as médias de cercárias eliminadas por dia, foram de 32,6% e 79 ± 90, respectivamente, para B. tenagophila de 11,3% e 112±100 para B. straminea e de 75,3% e 432 ± 436 para B. glabrata. As taxas mais baixas de infecção e as médias menores de cercárias eliminadas por B. tenagophila e B. straminea foram relacionadas aos sistemas internos de defesa mais eficientes dessas espécies.

A comparative study of the development of Schistosoma mansoni during the intra-molluscan phase was made by means of histological sections of Biomphalaria tenagophila, B. straminea and B. glabrata from Brazil. Two hundred snails of each species were individually exposed to 50 miracidia of the S. mansoni, AL line. No larvae were observed in the snails fixed 72 h after exposure. In specimens shedding cercariae, 31 days after exposure tissue reactions encapsulating the larvae were seen in B. tenagophila and B. straminea, in the head-foot, mantle collar and renal ducts. No tissue reactions occurred in the digestive glands of these two species. In B. glabrata the presence of numerous sporocysts and cercariae without tissue reactions was observed in the digestive gland, and other organs. The levels of infection of the snails and the average numbers of cercariae shed per day were 32.6% and 79±90 respectively for B. tenagophila, 11.3% and 112±100 for B. straminea and 75.3% and 432±436 for B. glabrata. The lower levels of infection and average numbers of cercariae shed by B. tenagophila and B. straminea are thus related to their more potent internal defense systems.

Aplicações trimestrais de niclosamida foram realizadas catorze anos em focos de Biomphalaria glabrata nas coleções hídricas de Peri-Peri, (Capim Branco, MG). Anualmente, os residentes da área eram submetidos a um exame coproscópio (Kato-Katz) e os que eliminavam ovos de Schistosoma mansoni nas fezes eram tratados com oxarnniquine. O levantamento malacológico trimestral foi realizado através de dez conchadas a cada dez metros nas duas margens das valas e córregos da região. Onde eram encontrados caramujos aplicava-se o moluscicida pela técnica de gotejamento ou aspersão de suspensão aquosa da niclosamida a 3 ppm. O índice médio de caramujos eliminando cercarias na região era de 14,3%. Após as quatro primeiras aplicações, foi reduzido a 0,0% e mantido em torno de 1,5% durante o programa. Portanto, a possibilidade de transmissão da esquistossomose na área continuou, e a população de caramujos se refazia em até três meses após aplicação do moluscicida. Os resultados obtidos neste estudo não encorajam o uso contínuo de niclosamida como medida única de controle da esquistossomose, por não diminuir permanentemente a população de planorbideos e por não interromper a possibilidade de transmissão da endemia.

Applications of niclosamide at three-monthly intervals were undertaken for 14 years in foci of Biomphalaria glabrata in the water sources of Peri-Peri (Capim Branco, MG). All the residents of the area were submitted to an annual fecal examination (Kato/Katz) and those individuals eliminating Schistosoma mansoni eggs were treated with oxamniquine. A malacological survey was undertaken at three-monthly intervals by means of ten scoops with a perforated ladle each ten metres along the two banks of the ditches and streams of the region. Where snails were found, molluscicide was applied by means of dripping or aspersion using a 3 ppm aqueous suspension of niclosamide. Initially, a mean of 14.3% of snails in the region were found to be eliminating cercariae. Following the first four applications of molluscicide, this was reduced to 0.0% and maintained at about 1.5% throughout the program. Thus, there was a continued possibility of schistosomiasis transmission in the area and it was observed that the population of snails reestablished itself within three months of molluscicide application. The results obtained in this study do not encourage the continual use of niclosamide as the only method of control of schistosomiasis.

Camundongos infectados experimentalmente com Schistosoma mansoni foram tratados por via oral com dose única de 125 ou 250 mg/kg de oltipraz, 50 ou 100 mg/kg de oxamniquine, e 200 ou 400 mg/kg de praziquantel. O número de vermes e alteração do oograma foram determinados entre a 1ª e 16ª semanas após o tratamento. O tempo necessário para observar o máximo de atividade da droga foi de 1 semana para o praziquantel, 2 semanas para o oxamniquine e 8 semanas para o oltipraz. Alterações do oograma persistiram durante o período de observação, embora recidiva tenha sido detectada, já na 4ª semana, com as drogas utilizadas.

Mice infected with adult Schistosoma mansoni were dosed with a single oral dose of 125 or 250 mg/kg oltipraz, 50 or 100 mg/kg oxamniquine, or 200 or 400 mg/kg praziquantel. The mortality rate of worms and oogram changes were determined between 1 and 16 weeks after dosing. The time required between dosing and postmortem to obtain maximum effectiveness was 1 week for praziquantel, 2 weeks for oxamniquine and 8 weeks for oltipraz. Changes in oograms persisted throughout most of the experiment, although relapse has been observed at the 4th week on.