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[Schistosomiasis in hyperendemic area of Taquarendi: I- Schistosoma mansoni infection and severe clinical forms].

The study was carried out in Taquarendi (Bahia), a caatinga zone with a small irrigated strip of land, where Biomphalaria glabrata snails are found. From the 1,532 inhabitants, 1,105 (72.1%) were submitted to clinical examination and 1058 out of them (95.7%) made stool examinations. Prevalence of schistosomiasis was 73.1% and 16.2% of these patients eliminated more than 1,000 eggs per gram of stool. By clinical examination, the size and the consistency of the left liver lobe were increased in 54% of the individuals and the spleen was palpable in 21.8%. The diagnosis of hepatosplenomegaly and of the advanced hepatointestinal clinical form was made, respectively, in 9.8% and 3.7%. A direct relationship between such clinical forms of the disease and the worm load over 1.000 S. mansoni eggs/g of fezes was found.

Adolescent↗

[A stock of infected snails for the mass breeding of cercariae and schistosomules].

A routine to maintain infected snails Biomphalaria glabrata able to produce 1-2 million cercariae per week and a mean shedding of about 3,500 cercariae per snail was quantitatively described. To a monthly production of about 5.3 and 6.5 million cercariae during the years of 1982 and 1983 it was necessary to expose an average of 1,557 and 1,957 snails, respectively. The efficiency described was maintained by infecting 2,000 snails per month (infectivity index of about 60%). A maximal production of 6,000 cercariae/snail was obtained in the period between 56-70 days after the snail infection by miracidia. By this period, however, 90% of the infected snails were dead. In the summer of 1982-1983 it was observed an impairment on the daily cercarial harvests which was related to ambient temperature increase, and the presence of rotifers in the aquaria water. When the cercariae were transformed into schistosomula in vitro, a yield of 55% schistosomula was obtained, with 7% of tail contamination. Lyophilization of cercariae produced 50.9 +/- 63 g dry weight per 1,000 cercariae.

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Latex of "coroa de cristo" (Euphorbia splendens): an effective molluscicide.

An aqueous solution of the latex of "coroa de cristo" (Euphorbia splendens var. hislopii) showed molluscicide action (LD90) at a concentration lower than 0.5 ppm on Biomphalaria glabrata and B. tenagophila reared in laboratory and at a concentration lower than 4.0 ppm for field B. tenagophila.

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[Occurrence of pleistocenic planorbids in the municipality of Jacobina, Bahia].

We registered Biomphalaria aff. glabrata (Say, 1818) from upper Pleistocene (or Holocene) based on paleontologic and stratigraphic data and in agreement with shell morphology. The shells came from Gruta das Onças, district of Caatinga do Moura, Jacobina country, state of Bahia. We also registered one more evidence of the ancient presence of water in the cave, which agree Cartelle & Bohorquez's (1982) hypothesis about the site.

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Drug resistance in schistosomiasis: a review.

Drug resistance associated with the treatment of human schistosomiasis appears to be an emerging problem requiring more attention from the scientific community than the subject currently receives. Drug-resistant strains of Schistosoma mansoni have been isolated by various investigators as a result of laboratory experimentation or from a combination of field and laboratory studies. Review of this data appears to indicate that the lack of susceptibility observed for some of the isolated strains cannot be ascribed solely to previous administration of antischistosome drugs and thus further studies are required to elucidate this phenomena. Strains of S. mansoni have now been identified from Brazil which are resistant to oxamniquine, hycanthone and niridazole; from Puerto Rico which are resistant to hycanthone and oxamniquine; and from Kenya which are resistant to niridazole and probably oxamniquine. Strains derived by in vitro selection and resistant to oxamniquine and possibly to oltipraz are also available. All of these strains are currently maintained in the laboratory in snails and mice, thus providing for the first time an opportunity for indepth comparative studies. Preliminary data indicates that S. haematobium strains resistant to metrifonate may be occurring in Kenya. This problem could poise great difficulty in the eventual development of antischistosomal agents. Biomphalaria glabrata from Puerto Rico and Brazil were found to be susceptible to drug-resistant S. mansoni from each country.

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[Isolation of a human strain of Schistosoma mansoni, Sambon,1907].

In this paper the authors briefly describe a human Schistosoma mansoni strain from Pará State, Brazil. The CIRENE'S strain was capable of infecting 71.4% of the snail vector Biomphalaria glabrata (Telegrafo's strain) provided by the "Evando Chagas" Institute, Belém. The cycle was completed by the infection of six mice. The thoracic and abdominal organs were examined microscopically which demonstrated the passage of the worm into the liver and lungs. The authors discuss the importance of these results in the epidemiology of schistosomiasis in Pará.

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[Molluscacide activity of piquerol A isolated from Piqueria trinervia (Compositae) on 8 species of pulmonate snails].

In laboratory trials an aqueous solution of Piquerol A from Piqueria trinervia, collected in several regions of Mexico, showed a molluscicide action on the adults of eight different pulmonates snails species: Fossaria (Fossaria) humilis, F. (Bakerilymnaea) sp., Pseudosuccinea columella and Stagnicola attenuata from Mexico; F. (B.) cubensis and Physa cubensis from Cuba; P. columella and Biomphalaria glabrata from Brazil; B. glabrata from Puerto Rico; and S. elodes from U.S.A. The solution was tested at 50, 25 and 5 ppm concentration, for two periods of 6 and 24 hours, at room temperature (20-22 degrees C). A 100% mortality was obtained for all species at 50 ppm concentration after 6 hours of exposure; the same percentage at 25 ppm after 24 hours; and 60 to 100% mortality at 5 ppm concentration during 24 hours of exposure. No recovery was observed among any of the treated snails. Piquerol A is a sesquiterpene with low stability in nature and has previously only been tested as an insecticide and as an inhibitor of metabolism in cell cultures: no field trails have been made on its toxicity to other aquatic fauna as yet, but it is believed Piquerol A could be an excellent molluscicide for use in areas where focal transmission of schistosomiasis and fascioliasis are taking place. This is the first time experiments on molluscicides have been carried out in Mexico.

Animals↗

Evaluation of the molluscicidal properties of Euphorbia splendens var. hislopii (N. E. B.) (Euphorbiaceae)--1. Experimental test in a lentic habitat.

The latex of Euphorbia splendens var. hislopii, at concentrations between 5 to 12 mg/l, kills 100% of the population of Biomphalaria glabrata in a lentic habitat, after 24 h. The lyophilized latex, stocked for 18 months, killed only 34.2% of the snails, at the concentration of 5 mg/l, and 96.0% at 12 mg/l. No lethal effect was observed among Pomacea haustrum exposed to the same concentrations of the molluscicide.

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Natural infection of wild rodents by Schistosoma mansoni. Parasitological aspects.

The evaluation of the role of rodents as natural hosts of Schistosoma mansoni was studied at the Pamparrão Valley, Sumidouro, RJ, with monthly captures and examination of the animals. Twenty-three Nectomys squamipes and 9 Akodon arviculoides with a schistosomal infection rate of 56.5% and 22.2% respectively eliminated a great majority of viable eggs. With a strain isolated from one of the naturally infected N. squamipes, we infected 75% of simpatric Biomphalaria glabrata and in 100% of albino Mus musculus mice. The adult worms, isolated from N. squamipes after perfusion were located mainly in the liver (91.5%) and the mesenteric veins (8.5%). The male/female proportion was of 2:1. The eggs were distributed on small intestine segments (proximal, medial and distal portions) and the large intestine without any significant differences in egg concentration of these segments. In A. arviculoides, the few eggs eliminated by the stools were viable and there was little egg retention on intestinal segments. Considering the ease to complete S. mansoni biological cycle in the Nectomys/Biomphalaria/Nectomys system under laboratory conditions, probably the same is likely to occur in natural conditions. In support to this hypothesis there are also the facts that human mansonic schistosomiasis has a very low prevalence in Sumidouro and endemicity among the rodents has not changed even after repeated treatments of the local patients. Based on our experiments, we conclude that N. squamipes has become a natural host of S. mansoni and possibly may participate in keeping the cycle of schistosomiasis transmission at Pamparrão Valley.

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[Control of schistosomiasis mansoni in an area of low transmission].

The schistosomiasis is transmitted by Biomphalaria tenagophila in our study area (Pedro de Toledo, São Paulo, Brazil). From 1980 to 1990 epidemiological surveys in a population of 4,000 inhabitants, has shown that: prevalences by Kato-Katz (KKT), immunofluorescence (FT) and intradermal (IDT) techniques were 22.8%, 55.5% and 51.8%, respectively; intensity of infection was low, 58.5 eggs per gram of faeces (epg); there were no symptomatic cases; prevalences were higher in mates, children and rural zone; index of potential contamination was 57.5% in the age group 5 to 20 years; 2/3 of patients were autochthonous; cases were no-randomly aggregated; transmission was focal and only 0.4% of snails were infected; water contacts through recreation showed the most important odds ratio; knowledge, attitudes and practices were satisfactory. From the epidemiological findings a control programme was carried out: yearly faeces exams, chemotherapy, molluscocide, health education and sanitation. Thus, the prevalence decreased sharply to 3.3% and intensity of infection to 30.3 epg; the incidence rates ranged between 0.4% and 2.5% annually; the sanitation became better and the youngsters were the main target in prophylaxis. To improve control, immunodiagnosis has to be conducted and the involvement of the population should be increase. However, we cannot forget that re-infection, therapeutic failure, etc, could play a major role in the maintenance this residual prevalence.

Adolescent↗

Identification of schistosome-infected snails by detecting schistosomal antigens and DNA sequences.

Cercarial shedding tests do not provide species identification of the schistosomes concerned and cannot detect prepatent schistosomal infections. We have demonstrated that both immunodetection by ELISA of schistosomal antigens in snail hemolymph, and dot hybridization of snail extracts by a DNA probe representing highly repeated sequences, proved suitable for detecting infected snails during prepatency as well as patency. A group-specific monoclonal antibody was found to be suitable for detecting Schistosoma mansoni infection in Biomphalaria sp., but not for positive identification of S. haematobium in Bulinus sp. Comparative evaluation of the diagnostic qualities, and technical aspects and cost of these tests, point to the superiority of the immunodetection approach for large scale detection of snails prepatently infected with S. mansoni. This approach is potentially useful for providing extended information on schistosome-snail epidemiology that may facilitate rapid evaluation of the danger of post-control reinfection, and help make decisions on the time and place of supplementary control measures. In this context the potential usefulness of the immunodetection or DNA probing approach for facilitating catalytic model representation of schistosome-snail epidemiology warrants further evaluation. Specific identification of S. haematobium in Bulinus by either of these approaches may be possible depending on the development of suitable antibodies or DNA probes.

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Intermediate hosts of Schistosoma mansoni in Brazil.

The Brazilian planorbidical chart is slowly but progressively been increased by new data. Distribution of vector species of Schistosoma mansoni, according to Paraense, 1986, may be thus resumed: Biomphalaria glabrata--delimited by parallels 13 and 21 degrees S and meridians 39 and 45 degrees W, area of greater dominance (Southeast Bahia, oriental hal of Minas Gerais and Espírito Santo). It is observed along the coast line of the states of Sergipe, Alagoas, Pernambuco, Paraiba and Rio Grande do Norte. Starting from there, it is found towards the southwest, in the direction to the São Francisco River and South-Center of Minas Gerais. Isolated population may be observed in other states. Its presence is probably, associated to the transmission of schistosomiasis in all areas where it occurs. B. tenagophila--extends it self through a wide strip of coast-line from the South of Bahia (17 degrees 45'S; 39 degrees 15'W), RS (33 degrees 41'S, 53 degrees 27'W). In São Paulo and Rio Grande do Sul states it is found further inland. It is important in schistosomiasis transmission in the Paraiba valley (SP). Isolated populations are observed in the Federal District and Minas Gerais state. B. straminea--better adapter species to climatic variation, having a more dense distribution in the northeast (41 degrees W and 110 degrees S), south of Bahia and northeast of Minas Gerais (150 and 180 degrees S, 400 and 440 degrees W). It is less susceptible than the B. glabrata, being however, the most important responsible for the transmission of S. mansoni in the northeast, chiefly in the northeastern dry area, where it is almost the only transmissive species.

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Schistosomiasis mansoni in the region of the Triângulo Mineiro, State of Minas Gerais, Brazil.

In order to reevaluate the possible presence of schistosomiasis mansoni in the Triângulo Mineiro, one of the areas of the State of Minas Gerais where this parasite is not commonly found, malacological survey and fecal examinations were undertaken in the region between October 1990 and June 1992. A sample of 7,032 1st grade school children from 29 counties had their feces examined using the Kato-Katz method. Amongst the children examined, two from Planura and one from each countie of Capinópolis, Conceição das Alagoas, Uberaba, Uberlândia, Prata and Gurinhatã were positive for Schistosoma mansoni. None of the children were identified as being autoctonous cases. In the malacological survey, 5,406 planorbid snails were examined. The specimens were identified morphologically and examined for S. mansoni by squashing between glass plates. The species were identified as Biomphalaria tenagophila in three counties, as B. straminea in ten and B. intermedia in 16. No snails were found in eight other counties studies. The snails were found to be negative for S. mansoni. The presence of intermediate hosts for S. mansoni, associated with parasitized individuals emphasizes the necessity of epidemiological surveillance for schistosomiasis in the region of Triângulo in the State of Minas Gerais.

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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.

Animals↗

DNA polymorphism of schistosomes and their snail hosts.

Analysis of the genomes of schistosomes and one of their intermediate hosts, Biomphalaria glabrata, using Random Amplified Polymorphic DNA (RAPD) demonstrated that intraspecific genetic polymorphism in the parasite is limited but in the snail is highly pronounced. This suggests an important role for the snail in the determination of the epidemiology of the disease. In addition to their intraspecific stability, schistosome derived RAPDs exhibit a high level of interspecific polymorphism and are thus ideal for the construction of phylogenetic trees. For the detection of intraspecific polymorphisms extensive variation in the mitochondrial DNA is being exploited for the development of a PCR based test for Schistosoma mansoni. Gene level polymorphisms are being analyzed by Low Stringency Single Specific Primer PCR.

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Current advances on the study of snail-snail interactions, with special emphasis on competition process.

Field work research on population dynamic of snails from the regions of Belo Horizonte and Lagoa Santa give much information about interactions among two or more species of mollusks: Pomacea haustrum, Biomphalaria glabrata, B. tenagophila, B. straminea and Melanoides tuberculata. Data ranging from two years to several decades ago suggest that the Pampulha reservoir is like a cemetery of B. glabrata and B. straminea, species that coexist for more than 14 years in a small part of a stream, whereas only B. glabrata lives in all the streams of the basin. In the last ten to twenty years B. tenagophila has coexisted with P. haustrum and M. tuberculata in the Serra Verde ponds and in the Pampulha dam. However these species have not settled in any of the brooks, except temporarily. The data suggest that the kind of biotope and the habitat conditions are decisive factors for the permanence of each species in its preferencial biotope. B. glabrata, natural from streams and riverheads, quickly disappears from the reservoirs and ponds where it coexists with other species for a short time, independently of the competitive process. Competition needs to be better studied, since in Central America and Caribean islands this kind of study has favored the biological control of planorbid species.

Animals↗

A laboratory-based approach to biological control of snails.

Development of Schistosoma mansoni in the intermediate host Biomphalaria glabrata is influenced by a number of parasite and snail genes. Understanding the genetics involved in this complex host/ parasite relationship may lead to an often discussed approach of introducing resistant B. glabrata into the field as a means of biological control for the parasite. For the snail, juvenile susceptibility to the parasite is controlled by at least four genes, whereas one gene seems to be responsible for adult nonsusceptibility. Obtaining DNA from F2 progeny snails from crosses between parasite-resistant and -susceptible snails, we have searched for molecular markers that show linkage to either the resistant or susceptible phenotype. Both restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) approaches have been used. To date, using a variety of snail and heterologous species probes, no RFLP marker has been found that segregates with either the resistant or susceptible phenotype in F2 progeny snails. More promising results however have been found with the RAPD approach, where a 1.3 kb marker appears in nearly all resistant progeny, and a 1.1 kb marker appears in all susceptible progeny.

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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.

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