Resumo Objetivo: descrever a distribuição geográfica dos hospedeiros intermediários do Schistosoma mansoni em cinco estados brasileiros. Métodos: estudo transversal; foram selecionados municípios dos estados do Paraná (78), Minas Gerais (120), Bahia (82), Pernambuco (51) e Rio Grande do Norte (98), nos anos de 2012 a 2014; esses municípios foram escolhidos por não possuírem registros atualizados da presença de caramujos hospedeiros intermediários de S. mansoni; moluscos foram capturados, taxonomicamente identificados e examinados para verificação de cercárias de S. mansoni. Resultados: os trabalhos foram realizados em 427 municípios (99,5% dos 429 selecionados); foi registrada presença de moluscos em 300 (70,2%) municípios e a ocorrência de Biomphalaria glabrata em 62 (21%) municípios, B. straminea em 181 (60%), B. tenagophila em três (1%); associação de B. glabrata/B. straminea foi encontrada em 53 (18%), e de B. glabrata/B. tenagophila em um (0,3%). Conclusão: os registros de B. glabrata, B. straminea e B. tenagophila estão de acordo com a distribuição conhecida.

Abstract Objective: to describe the geographical distribution of intermediate hosts of Schistosoma mansoni in five Brazilian states. Methods: this was a descriptive cross-sectional study; municipalities were selected in the states of Paraná (78), Minas Gerais (120), Bahia (82), Pernambuco (51) , and Rio Grande do Norte (98), for the period 2012 to 2014; these municipalities were chosen because they did not have current records of the presence of snails vectores de S. mansoni. The molluscs were captured and taxonomically identified and examined for S. mansoni cercariae. Results: the work was carried out in 427 municipalities (99.5% of the 429 selected); the presence of mollusks was registered in 300 (70.2%) municipalities; Biomphalaria glabrata were found in 62 (21%) municipalities, B. straminea in 181 (60%), B. tenagophila in three (1%); B. glabrata/B. straminea association was found in 53 municipalities (18%) and B. glabrata/B. tenagophila association in one (0.3%) municipality. Conclusion: B. glabrata, B. straminea and B. tenagophila distribution records obtained in this study are consistent with previously known distribution.

Resumen Objetivo: describir la distribución geográfica de los hospedadores intermediarios de Schistosoma mansoni en cinco estados brasileños. Métodos: estudio epidemiológico transversal; el estudio fue realizado en municipios de los estados de Paraná (78), Minas Gerais (120), Bahia (82), Pernambuco (51) y Rio Grande do Norte (98), entre 2012 y 2014; estos municipios fueron elegidos por no poseer registros actualizados de la presencia de caracoles vectores de S. mansoni; los moluscos fueron capturados, taxonómicamente identificados y examinados para la verificación de cercarias de S. mansoni. Resultados: los trabajos fueron realizados en 427 municipios (99,5% de 429 municipios seleccionados); fue registrada presencia de moluscos en 300 (70,2%) municípios; la presencia de Biomphalaria glabrata fue registrada en 62 (21%) municipios, B. straminea en 181 (60%) y B. tenagophila en três (1%); se observó asociación de B. glabrata con B. straminea en 53 (18%) y de B. glabrata con B. tenagophila en uno (0,3%). Conclusión: los registros de Biomphalaria están de acuerdo con la distribución conocida.

Resumo Objetivo: descrever a distribuição geográfica dos hospedeiros intermediários do Schistosoma mansoni em cinco estados brasileiros. Métodos: estudo transversal; foram selecionados municípios dos estados do Paraná (78), Minas Gerais (120), Bahia (82), Pernambuco (51) e Rio Grande do Norte (98), nos anos de 2012 a 2014; esses municípios foram escolhidos por não possuírem registros atualizados da presença de caramujos hospedeiros intermediários de S. mansoni; moluscos foram capturados, taxonomicamente identificados e examinados para verificação de cercárias de S. mansoni. Resultados: os trabalhos foram realizados em 427 municípios (99,5% dos 429 selecionados); foi registrada presença de moluscos em 300 (70,2%) municípios e a ocorrência de Biomphalaria glabrata em 62 (21%) municípios, B. straminea em 181 (60%), B. tenagophila em três (1%); associação de B. glabrata/B. straminea foi encontrada em 53 (18%), e de B. glabrata/B. tenagophila em um (0,3%). Conclusão: os registros de B. glabrata, B. straminea e B. tenagophila estão de acordo com a distribuição conhecida.

Abstract Objective: to describe the geographical distribution of intermediate hosts of Schistosoma mansoni in five Brazilian states. Methods: this was a descriptive cross-sectional study; municipalities were selected in the states of Paraná (78), Minas Gerais (120), Bahia (82), Pernambuco (51) , and Rio Grande do Norte (98), for the period 2012 to 2014; these municipalities were chosen because they did not have current records of the presence of snails vectores de S. mansoni. The molluscs were captured and taxonomically identified and examined for S. mansoni cercariae. Results: the work was carried out in 427 municipalities (99.5% of the 429 selected); the presence of mollusks was registered in 300 (70.2%) municipalities; Biomphalaria glabrata were found in 62 (21%) municipalities, B. straminea in 181 (60%), B. tenagophila in three (1%); B. glabrata/B. straminea association was found in 53 municipalities (18%) and B. glabrata/B. tenagophila association in one (0.3%) municipality. Conclusion: B. glabrata, B. straminea and B. tenagophila distribution records obtained in this study are consistent with previously known distribution.

Resumen Objetivo: describir la distribución geográfica de los hospedadores intermediarios de Schistosoma mansoni en cinco estados brasileños. Métodos: estudio epidemiológico transversal; el estudio fue realizado en municipios de los estados de Paraná (78), Minas Gerais (120), Bahia (82), Pernambuco (51) y Rio Grande do Norte (98), entre 2012 y 2014; estos municipios fueron elegidos por no poseer registros actualizados de la presencia de caracoles vectores de S. mansoni; los moluscos fueron capturados, taxonómicamente identificados y examinados para la verificación de cercarias de S. mansoni. Resultados: los trabajos fueron realizados en 427 municipios (99,5% de 429 municipios seleccionados); fue registrada presencia de moluscos en 300 (70,2%) municípios; la presencia de Biomphalaria glabrata fue registrada en 62 (21%) municipios, B. straminea en 181 (60%) y B. tenagophila en três (1%); se observó asociación de B. glabrata con B. straminea en 53 (18%) y de B. glabrata con B. tenagophila en uno (0,3%). Conclusión: los registros de Biomphalaria están de acuerdo con la distribución conocida.

Angiostrongylus cantonensis is the most common aetiological agent of human eosinophilic meningoencephalitis. Following a report indicating the presence of this parasite in Brazil in 2007, the present study was undertaken to investigate the presence of A. cantonensis in the surrounding Brazilian port areas. In total, 30 ports were investigated and the following molluscs were identified: Achatina fulica, Belocaulus sp., Bradybaena similaris sp., Cyclodontina sp., Helix sp., Leptinaria sp., Melampus sp., Melanoides tuberculata, Phyllocaulis sp., Pomacea sp., Pseudoxychona sp., Rhinus sp., Sarasinula marginata, Streptaxis sp., Subulina octona, Succinea sp., Tomigerus sp., Wayampia sp. and specimens belonging to Limacidae and Orthalicinae. Digestion and sedimentation processes were performed and the sediments were examined. DNA was extracted from the obtained larvae and the internal transcribed spacer region 2 was analysed by polymerase chain reaction-restriction fragment length polymorphism after digestion with the endonuclease ClaI. Of the 30 ports investigated in this study, 11 contained molluscs infected with A. cantonensis larvae. The set of infected species consisted of S. octona, S. marginata, A. fulica and B. similaris. A total of 36.6% of the investigated ports were positive for A. cantonensis, indicating a wide distribution of this worm. It remains uncertain when and how A. cantonensis was introduced into South America.

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.

INTRODUÇÃO: O aumento das práticas de ecoturismo e turismo rural, em Minas Gerais, Brasil, evidencia a importância de se realizarem estudos sobre a ocorrência de hospedeiros intermediários do Schistosoma mansoni, no estado. O presente trabalho objetivou a busca e identificação das espécies de caramujos Biomphalaria encontrados em municípios mineiros pertencentes à Estrada Real, um importante projeto de turismo brasileiro. MÉTODOS: Os moluscos foram coletados em 36 municípios da Estrada Real, no sudeste de Minas Gerais. A pesquisa foi realizada de agosto de 2005 a setembro de 2009 e todos os locais visitados foram georreferenciados com o uso de GPS. RESULTADOS: Dos 36 municípios estudados, 30 apresentaram a ocorrência de pelo menos uma entre as seis espécies de Biomphalaria: glabrata, tenagophila, straminea, peregrina, occidentalis e schrammi. As três primeiras espécies citadas, reconhecidas como hospedeiras intermediárias do S. mansoni, estavam presentes em 33,3%, 47,2% e 8,3% dos municípios estudados, respectivamente. Os moluscos foram encontrados em diferentes tipos de coleções hídricas e em nenhum deles foi detectada infecção pelo S. mansoni. Houve maior ocorrência de Biomphalaria na área referente ao Caminho Novo (Diamantina/MG ao Rio de Janeiro/RJ). CONCLUSÕES: Considerando-se a ocorrência da esquistossomose, no Estado de Minas Gerais, e as repercussões socioeconômicas que envolvem o projeto Estrada Real, este trabalho aponta para a vulnerabilidade das coleções hídricas devido à presença de moluscos Biomphalaria e enfatiza a necessidade de vigilância epidemiológica e medidas educativas e sanitárias integradas com a comunidade local e setores de turismo.

INTRODUCTION: The increasing practice of ecotourism and rural tourism in the State of Minas Gerais, Brazil, highlights the importance of studies concerning the occurrence of potential intermediate hosts of Schistosoma mansoni. This study aimed to identify species of Biomphalaria snails in municipalities along the Estrada Real, an important Brazilian tourism project. METHODS: The specimens were collected in different water collections of 36 municipalities along the Estrada Real in the southeast of the State of Minas Gerais. Biomphalaria species were characterized using both morphological and molecular approaches. The research was conducted between August 2005 and September 2009 and all the sites visited were georeferenced using GPS. RESULTS: Six Biomphalaria species were found in 30 of the 36 municipalities studied: glabrata, tenagophila, straminea, peregrina, occidentalis and schrammi. The first three species of Biomphalaria, recognized as intermediate hosts of S. mansoni, were present in 33.3%, 47.2% and 8.3% of the municipalities studied, respectively. The mollusks were found in different types of water collections and no infection by S. mansoni was detected. The highest occurrence of Biomphalaria concentration was verified in the area covered by the Caminho Novo route (Diamantina/MG to Rio de Janeiro/RJ). CONCLUSIONS: Considering the occurrence of schistosomiasis in the State of Minas Gerais and the socioeconomic repercussions involved in the Estrada Real Project, this work focuses on the vulnerability of water collections due to the presence of Biomphalaria mollusks and emphasizes the need for epidemiological surveillance and sanitary and educational measures integrated with the local community and tourism sectors.

Mitochondrial DNA of Biomphalaria tenagophila, a mollusc intermediate host of Schistosoma mansoni in Brazil, was sequenced and characterised. The genome size found for B. tenagophila was 13,722 bp and contained 13 messenger RNAs, 22 transfer RNAs (tRNA) and two ribosomal RNAs (rRNA). In addition to sequencing, the mitochondrial DNA (mtDNA) genome organization of B. tenagophila was analysed based on its content and localization of both coding and non-coding regions, regions of gene overlap and tRNA nucleotide sequences. Sequences of protein, rRNA 12S and rRNA 16S nucleotides as well as gene organization were compared between B. tenagophila and Biomphalaria glabrata, as the latter is the most important S. mansoni intermediate host in Brazil. Differences between such species were observed regarding rRNA composition. The complete sequence of the B. tenagophila mitochondrial genome was deposited in GenBank (accession EF433576). Furthermore, phylogenetic relationships were estimated among 28 mollusc species, which had their complete mitochondrial genome deposited in GenBank, using the neighbour-joining method, maximum parsimony and maximum likelihood bootstrap. B. tenagophila was positioned at a branch close to B. glabrata and Pulmonata molluscs, collectively comprising a paraphyletic group, contrary to Opistobranchia, which was positioned at a single branch and constituted a monophyletic group.

Plantas consumidas como remédio nem sempre são identificadas taxonomicamente de maneira correta. Se estas plantas forem utilizadas para obtenção de uma droga vegetal ou um fitoterápico, tal erro pode afetar a qualidade do produto final. Neste trabalho são descritos marcadores para a identificação de Solanum cernuum Vell. (Solanaceae), esteja a planta íntegra, triturada ou como extrato bruto. Indivíduos de quatro localidades de Minas Gerais foram coletados, analisados e comparados. Os caracteres morfológicos foram utilizados para a planta íntegra. Para a planta triturada, o conjunto dos tricomas foi utilizado como marcador anatômico. Um marcador macromolecular também foi determinado. Para tal a região ITS1, 5.8S e ITS2 do DNAr foi clonada e seqüenciada. A seqüência, com cerca de 600 pares de bases dos quais 48,1% são AT, foi depositada no GenBank sob o número de acesso DQ837371. Por ser uma seqüência específica para S. cernuum, ela pode ser usada como marcador desta espécie. Para o extrato bruto foram determinados perfis cromatográficos de extratos das folhas por cromatografia em camada delgada e por cromatografia líquida de alta eficiência. Dois flavonóides foram isolados e identificados como quercitrina e afzelina. Assim, neste trabalho foram determinados marcadores morfológicos, anatômicos, macro e micromoleculares para identificar S. cernuum.

The plants consumed as remedy by the population may have imprecise taxonomical identification. If these plants are used for the production of phytomedicines such misidentification may affect the quality of the product. Hereby, we describe markers for identification of the entire plant or grounded plant material or the crude extract of Solanum cernuum Vell. (Solanaceae). Specimens from four localities were collected, analyzed and compared. Morphological characters were used to identify the plant when it is not grounded or extracted. However, when the plant material is grounded, the set of trichomes may be used as anatomical marker. The region ITS1, 5.8S and ITS2 of the nuclear ribosomal DNA was cloned and sequenced. The sequence, with length of about 600 base pairs, being 48.1% AT , was deposited in GenBank under the accession number DQ837371. Once this sequence is specific to S. cernuum, it was used as marker for this species. For the crude extract, chromatographic profiles of the leaves extracts were obtained by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). Two flavonoids were isolated and identified as quercitrin and afzelin. So, this study presents morphological, anatomical, macro and micromolecular markers to identify S. cernuum.

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 study focuses on the geographic distribution of the snail of the genus Biomphalaria and evaluates its infectivity by Schistosoma mansoni in 5264 specimens collected in the municipality of Juiz de Fora, Minas Gerais, Brazil. Of the 31 locations studied, 6 were reservoirs, 11 rudimentary holding ponds, 7 irrigation ditches, 5 lakes, 1 ornamental pond, and 1 waterfall. Intermediate hosts were found only in the rudimentary ponds and ditches, which were 100% positive. Using morphological and molecular analysis techniques, B. tenagophila, B. peregrina, and B. straminea were identified. This is the first report of B. stramínea in the municipality, and evaluation of its infective potential revealed susceptibility of 25.4%. Although we did not find specimens of Biomphalaria infected by S. mansoni, the data obtained indicate the presence of intermediate hosts, especially in the irrigation ditches in Juiz de Fora, and their proximity to contaminated areas.

Jaboticatubas is a municipality in the metropolitan region of Belo Horizonte which has been a target of a wide media release as "the capital of schistosomiasis" since the 1960's. In order to give support to a work based on an integrated control, we sought to identify the disease determinants at the site. A transversal study was carried out aimed at identifying prevalence rates of the disease and factors associated with the infection in the district of São José de Almeida, and two close localities, Cipó Velho and São José da Serra, all of them located in the municipality of Jaboticatubas. A parasitological survey was performed, applying the Kato-Katz method with two slides per sample in 1186 schoolchildren which represents 77% of all registered pupils in four public schools in 2001. Among these schoolchildren a number of 101 (8.6%) prooved positive for Schistosoma mansoni eggs in their stool samples. A total of 64 families, whose schoolchildren had shown to be positive for schistosomiasis, also undertook examinations. As negative control, a random sample was collected from the 206 families, whose children had proven negative for schistosomiasis. The prevalence among 270 families (1304 people) was 12%. To assess those who continued to have contact with possibly contaminated water, 1061 (81.4%) people of the 270 families were interviewed. A multivariate analysis identified the following factors associated with the infection: time of residence in the area (short period), garbage disposal (use of deserted areas), gender (male), age (from 10 to 29 years), and water contact (daily and weekly). Further analysis of these factors revealed a close correlation between water contact and the disease, with a positive significant frequency concerning almost all those items. Depending on gender and age significant variations of water contact patterns associated with leisure and professional activities were found. A malacological survey on water collections in the area identified snails of the species Biomphalaria straminea and B. glabrata. The latter showed 17 (0.6%) specimens positive for S. mansoni. Qualitative studies have complemented such evidences, which allowed us to design a reference picture and specific indicators of the disease for the local population. Those data provided the essential information to continue the development of an already ongoing educative process, as well as projects on environmental improvements.

Recently, the booming rural tourism in endemic areas of the state of Minas Gerais was identified as a contributing factor in the dissemination of the infection with Schistosoma mansoni. This article presents data from six holiday resorts in a rural district approximately 100 km distant from Belo Horizonte, MG, Brazil, where a possibly new and until now unperceived way of transmission was observed. The infection takes place in swimming pools and little ponds, which are offered to tourists and the local population for fishing and leisure activities. The health authorities of the district reported cases of schistosomiasis among the local population after visiting these sites. As individuals of the non-immune middle class parts of the society of big urban centers also frequent these resorts, infection of these persons cannot be excluded. A malacological survey revealed the presence of molluscs of the species Biomphalaria glabrata and Biomphalaria straminea at the resorts. The snails (B. glabrata) of one resort tested positive for S. mansoni. In order to resolve this complex problem a multidisciplinary approach including health education, sanitation measures, assistance to the local health services, and evolvement of the local political authorities, the local community, the tourism association, and the owners of the leisure resorts is necessary. This evidence emphasizes the urgent need for a participative strategic plan to develop the local tourism in an organized and well-administered way. Only so this important source of income for the region can be ensured on the long term without disseminating the disease and putting the health of the visitors at risk.

Freshwater snails belonging to the genus Biomphalaria act as intermediate hosts for the parasite trematode Schistosoma mansoni in Africa and in the neotropical region. Identification of such molluscs is carried out based on morphological characters and the presence of cercariae is verified through squeezing snails between two glass slides or by exposing them to artificial light. However, sometimes, the material collected includes molluscs with decomposed bodies or, yet, only empty shells, which precludes their identification and S. mansoni detection. Due to these difficulties, we have developed a methodology in which DNA may be extracted from traces of organic material from inside shells in order to identify molluscs through polymerase chain reaction and restriction fragment length polymorphism and to detect S. mansoni into these snails, by using low stringency polymerase chain reaction. Species-specific profiles obtained from B. glabrata, B. straminea, and B. tenagophila snails and their shells, maintained in laboratory for ten years, showed the same profiles. S. mansoni profiles showed to be present in shell specimens as far as the eighth week after being removed from aquarium.

Published and unpublished observations on geographical distribution of Biomphalaria snails in the State of Minas Gerais, Brazil, were compiled. This work is aimed at knowing the present occurrence of Biomphalaria species in this region, and at contributing to the elaboration of the planorbid chart of Minas Gerais. In malacological surveys, performed by several researchers, the presence of seven species of this genus was recorded. Those planorbids were found in 12 mesoregions, in 283 (33.1%) municipalities out of 853 with the following distribution: B. glabrata (185 municipalities), B. straminea (125), B. tenagophila (58), B. peregrina (57), B. schrammi (26), B. intermedia (20) and B. occidentalis (2). B. glabrata and B. tenagophila are found naturally infected by Schistosoma mansoni in Minas Gerais. In 24 municipalities the three snail hosts of S. mansoni in Brazil, B. glabrata, B. tenagophila and B. straminea, are present.

O objetivo desse trabalho foi determinar a prevalência das parasitoses em escolares de Bambuí, através de exames coprológicos (direto e Kato-Katz) e reavaliar os criadouros de moluscos descritos no município. Dos 2.901 escolares examinados, 20,1% estavam parasitados, sendo que Giardia lamblia, Entamoeba coli, Ascaris lumbricoides e ancilostomídeos foram os parasitas mais freqüentes, com prevalências de 6,2%, 6,2%, 4,8% e 1,4%, respectivamente. Os ancilostomídeos foram significativamente mais freqüentes na zona rural e nos alunos com mais de 14 anos, enquanto a prevalência da E. coli foi maior na zona urbana e a G. lamblia mais freqüente na faixa etária de 0-6 anos. Somente três crianças eliminavam ovos de Schistosoma mansoni. O único hospedeiro intermediário encontrado foi a Biomphalaria glabrata e nenhuma delas estava eliminando cercárias de S. mansoni . Comparando-se estes dados a de levantamentos realizados anteriormente no município, observou-se uma queda na prevalência de todos os parasitas. Algumas hipóteses para tentar explicar esta queda são discutidas tais como: processo intenso de urbanização e a melhoria das condições sócio-sanitárias do município.

This work was carried out with the purpose of determining the prevalence of intestinal parasitoses in the school children of Bambuí, through parasitological examinations (direct and Kato-Katz methods) and reevaluating the snails' breeding places described in the county. Of the 2,091 school children examined, 20.1% had at least one parasitic infection. Giardia lamblia, Entamoeba coli, Ascaris lumbricoides and hookworm are the most frequent parasites, with a prevalence of 6.2%, 6.2%, 4.8% and 1.4%, respectively. The hookworms were significantly more frequent among students from the rural area and in those aged over 14 years, while the prevalence of E. coli was greater in the urban area and the G. lamblia was more frequent in children under 6 years old. Only three children shed eggs of Schistosoma mansoni. The only intermediate host found was Biomphalaria glabrata and none of them was shedding cercariae of S. mansoni. When these data were compared with data from other surveys previously effected in the county, a decline was observed in the prevalence of all parasites. Some hypotheses which may explain this decline are discussed, such as: intense urbanization process and improvement of social-sanitary conditions of the county.