Cecília Pereira de Souza

Origin and diversification of the human parasite Schistosoma mansoni

Schistosoma mansoni is the most widespread of the human‐infecting schistosomes, present in 54 countries, predominantly in Africa, but also in Madagascar, the Arabian Peninsula, and the Neotropics. Adult‐stage parasites that infect humans are also occasionally recovered from baboons, rodents, and other mammals. Larval stages of the parasite are dependent upon certain species of freshwater snails in the genus Biomphalaria, which largely determine the parasites geographical range. How S. mansoni genetic diversity is distributed geographically and among isolates using different hosts has never been examined with DNA sequence data. Here we describe the global phylogeography of S. mansoni using more than 2500 bp of mitochondrial DNA (mtDNA) from 143 parasites collected in 53 geographically widespread localities. Considerable within‐species mtDNA diversity was found, with 85 unique haplotypes grouping into five distinct lineages. Geographical separation, and not host use, appears to be the most important factor in the diversification of the parasite. East African specimens showed a remarkable amount of variation, comprising three clades and basal members of a fourth, strongly suggesting an East African origin for the parasite 0.30–0.43 million years ago, a time frame that follows the arrival of its snail host. Less but still substantial variation was found in the rest of Africa. A recent colonization of the New World is supported by finding only seven closely related New World haplotypes which have West African affinities. All Brazilian isolates have nearly identical mtDNA haplotypes, suggesting a founder effect from the establishment and spread of the parasite in this large country.

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.

Degree of host-parasite compatibility between Schistosoma mansoni and their intermediate molluscan hosts in Brazil

The compatibility of Biomphalaria tenagophila, B. straminea and B. glabrata from Minas Gerais with different strains of Schistosoma mansoni was evaluated using the method of Frandsen (1979b) in standardized experiments. One hundred and fifty of each species of snail were individually exposed in the laboratory to 50 miracidia of S. mansoni lines LE, SJ and AL. The cercariae from the infected snails were counted and used to calculate TCP/100 indices, which were compared with those of Frandsen (1979b). For B. tenagophila the TCP/100 indices varied from 37,996 to 74,266 (class II and III). The snail was poorly compatible with LE (class II) and compatible with SJ and AL (class III). For B. straminea the indices varied from 9,484 to 20,508. The snail was not very compatible with SJ (class I) and poorly compatible with LE and AL (class II). For B. glabrata the indices varied from 588,828 to 1,039,065. The snails was extremely compatible (class VI) with the three lines of S. mansoni. These results confirm the epidemiological importance of B. glabrata in Brazil followed by B. tenagophila and B. straminea.

The Distribution of Biomphalaria spp. in Different Habitats in Relation to Physical, Biological, Water Contact and Cognitive Factors in a Rural Area in Minas Gerais, Brazil

A total of 256 sites in 11 habitats were surveyed for Biomphalaria in Melquiades rural area (State of Minas Gerais) in August and November 1999 and in March 2000. Of the 1,780 Biomphalaria collected, 1,721 (96.7%) were B. glabrata and 59 (3.3%) B. straminea. Snails were found in all habitats except in wells, with the largest mean numbers in tanks, seepage ponds and canals, and the smallest numbers in springs, rice fields and fishponds. Peoples knowledge of the occurrence of Biomphalaria at the collection sites and the presence of Biomphalaria ova were strongly correlated with the occurrence of snails, and distance between houses and collection sites, as well as water velocity were inversely correlated with Biomphalaria occurrence (p < 0.001). The strongest predictor o f Biomphalaria occurrence was the presence of tilapia fish in fishponds. Fourteen Biomphalaria (0.8% of all snails) found at 6 sites were infected with Schistosoma mansoni. Suggestions are made for the utilization of local peoples knowledge in snail surveys and further studies are recommended on the possible use of tilapia for biological control of Biomphalaria in fishponds, as well as modeling of S. mansoni transmission and reinfection.

Moluscos límnicos da microrregião de Belo Horizonte, MG, com ênfase nos vetores de parasitoses

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.

DEVELOPMENT OF SCHISTOSOMA MANSONI IN BIOMPHALARIA TENAGOPHILA, BIOMPHALARIA STRAMINEA AND BIOMPHALARIA GLABRATA

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.

O uso da casca da castanha do caju, Anacardium occidentale, como moluscicida alternativo

Bioassays using hexanolic extracts of cashew nut shells, of Anacardium occidentale, collected in Ceara 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.

Ensaios preliminares em laboratório para verificar a ação moluscicida de algumas espécies da flora brasileira

Estudou-se em laboratorio a atividade moluscicida de 68 extratos de 23 plantas brasileiras. As solucoes em agua desclorada dos extratos hexânicos e etanolico, nas concentracoes de 1, 10 e 100 ppm, foram testadas sobre caramujos adultos e desovas de Biomphalaria glabrata, criados em laboratorio. As plantas que demonstraram acao moluscicida na concentracao de 100 ppm foram: Arthemisia verlotorum Lamotte, Caesalpinia peltophoroides Benth, Cassia rugosa G.Don., Eclipta alba Hassk, Euphorbia pulcherrima Willd, Euphorbia splendens Bojer, Joannesia princeps Vell, Leonorus sibiricus L.,Macrosiphonia guaranitica Muell,Nerium oleander L., Palicourea nicotianaefolia Cham, e Schlech., Panicum maximum M., Rumex crispus L., Ruta graveolens L., e Stryphnodendron barbatiman M.Estudou-se em laboratorio a atividade moluscicida de 68 extratos de 23 plantas brasileiras. As solucoes em agua desclorada dos extratos hexânicos e etanolico, nas concentracoes de 1, 10 e 100 ppm, foram testadas sobre caramujos adultos e desovas de Biomphalaria glabrata, criados em laboratorio. As plantas que demonstraram acao moluscicida na concentracao de 100 ppm foram: Arthemisia verlotorum Lamotte, Caesalpinia peltophoroides Benth, Cassia rugosa G.Don., Eclipta alba Hassk, Euphorbia pulcherrima Willd, Euphorbia splendens Bojer, Joannesia princeps Vell, Leonorus sibiricus L.,Macrosiphonia guaranitica Muell,Nerium oleander L., Palicourea nicotianaefolia Cham, e Schlech., Panicum maximum M., Rumex crispus L., Ruta graveolens L., e Stryphnodendron barbatiman M.

Aspects of the Maintenance of the Life Cycle of Fasciola hepatica in Lymnaea columella in Minas Gerais, Brazil

Fascioliasis is a parasitic disease of domestic ruminants that occurs worldwide. The lymnaeid intermediate hosts of Fasciola hepatica include Lymnaea columella, which is widely distributed in Brazil. A colony of L. columella from Belo Horizonte, MG, was reared in our laboratory to be used in studies of the F. hepatica life cycle, the intermediate host-parasite relationship and development of an anti-helminthic vaccine. In the first experiment 1,180 snails were exposed to miracidia of F. hepatica eggs removed from the biliary tracts of cattle from the State of Rio Grande do Sul. In the second and third experiments the snails were exposed to miracidia that had emerged from F. hepatica eggs from Uruguay, maintained in rabbits. The rates of infection in the first, second and third experiments were 0, 42.1 and 0% respectively. Over 15,806 metacercariae were obtained and stored at 4 degrees C. Four rabbits weighing 1.5 kg each were infected with 32-44 metacercariae and two with 200. Three rabbits begin to eliminate eggs of the parasite in the feces from 84 days after infection onwards. The biological cycle of F. hepatica in L. columella and the rabbit was completed within 124 days.

Molluscicide control of snail vectors of schistosomiasis

A review of the methodology recommended by the World Health Organization for the use of molluscicides for the control of snail vectors of schistosomiasis is presented. Discussion of the principle molluscicides used, their advantages and disadvantages, the techniques and equipment required for their application and evaluation of effect as well as the biological control of snails is included.