Participation of N-acetyl-D-glucosamine carbohydrate moieties in the recognition of Schistosoma mansoni sporocysts by haemocytes of Biomphalaria tenagophila

Lectin-carbohydrate binding may be involved in the recognition of Schistosoma mansoni sporocysts by haemocytes of Biomphalaria; therefore, we tested if this interaction is associated with snail resistance against Schistosoma infection. In vitro data showed that most of the S. mansoni sporocysts cultured with haemocytes from Biomphalaria glabrata BH, a highly susceptible snail strain, had a low number of cells that adhered to their tegument and a low mortality rate. Moreover, the addition of N-acetyl-D-glucosamine (GlcNAc) did not alter this pattern of adherence and mortality. Using haemocytes and haemolymph of Biomphalaria tenagophila Cabo Frio, we observed a high percentage of sporocysts with adherent cells, but complete encapsulation was not detected. Low concentrations of GlcNAc increased haemocyte binding to the sporocysts and mortality, which returned to basal levels with high concentrations of the carbohydrate. In contrast, haemocytes plus haemolymph from B. tenagophila Taim encapsulated cellular adhesion index of level 3 and destroyed over 30% of the S. mansoni sporocysts in culture. Interestingly, the addition of GlcNAc, but not mannose, to the culture medium resulted in the significant inhibition of cellular adhesion to the parasite tegument and the reduction of parasite mortality, suggesting that GlcNAc carbohydrate moieties are important to the recognition of S. mansoni by B. tenagophila Taim.

Changes in the behavioral and immunological parameters of the mollusk Biomphalaria tenagophila induced by disruption of the circadian cycle as a consequence of continuous illumination

In the present investigation we studied some behavioral and immunological parameters of adult gastropod mollusk, Biomphalaria tenagophila, which have been reproducing for several generations under laboratory conditions. One group of gastropods was kept on a 14-h light/10-h dark cycle, corresponding to a regular circadian cycle, and another group was exposed to continuous light for 48 h. Animals were studied along (behavioral groups) or immediately after (immunological groups) 48 h of regular circadian cycle or continuous light conditions. Stopping/floating, dragging and sliding were the behavioral aspects considered (N = 20 for regular cycle; N = 20 for continuous illumination) and number of hemocytes/µl hemolymph was the immunological parameter studied (N = 15 for regular cycle, N = 14 for continuous illumination). Animals under continuous illumination were more active (sliding = 33 episodes, dragging = 48 episodes) and displayed a lower number of hemocytes (78.0 ± 24.27/µl) when compared with mollusks kept on a regular circadian cycle (sliding = 18 episodes, dragging = 27 episodes; hemocytes = 157.6 ± 53.27/µl). The data are discussed in terms of neural circuits and neuroimmunological relations with the possible stressful effect of continuous illumination.

Biomphalaria tenagophila: genetic variability within intermediate snail hosts susceptible and resistant to Schistosoma mansoni infection

DNA analysis by molecular techniques has significantly expanded the perspectives of the study and understanding of genetic variability in molluscs that ere vectors of schistosomiasis. In tire present study, the genetic variability of susceptible and resistant B. tenagophila strains to S. mansoni infection was investigated using amplification of their genomic DNA by RAPD-PCR. The products were analyzed by PAGE and stained with silver. The results showed pdymorphism between tested strains with four different primers. We found two bonds of 1,900 and 3,420 bp that were characteristic of the susceptible strains with primer 2. The primers 9 end 10 identified a single polymorphic bond that was also characteristic of (3,136 and 5,041 bp, respectively) susceptible snails. Two polymorphic bonds were detected by primer 15: one with 1 800 bp was characteristic of the resistant strain and the other with 1,700 do in the susceptible one. These results provide additional evidence showing that the RAPD-PCR technique is adequate for the study of polymorphisms in intermediate hosts snails of S. mansoni. The obtained results are expected to expend the knowledge about the genetic variability of the snails and to permit the future identification of genomic sequences specifically related to the resistance/susceptibility of Biompholario to the larval forms of S. mansoni.

Differences in the number of hemocytes in the snail host Biomphalaria tenagophila, resistant and susceptible to Schistosoma mansoni infection

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Suscetibilidade de biomphalaria glabrata, B. straminea e B. tenagophila a diferentes cepas de schistosoma mansoni

Em condições experimentais foi estudada a suscetibilidade de Biomphalaria glabrata, B. straminea e B. tenagophila a quatro linhagens humanas (MAP, PTH, UPH, e OuH) e duas de roedores silvestres (PTR e VPR) do Schistosoma mansoni. Grupos de 50 moluscos foram expostos individualmente a 10 miracídios e observados durante 70 dias. Avaliou-se a suscetibilidade dos moluscos ao parasito por meio da % de animais com esporocistos, % de moluscos que eliminavam cercárias e mortalidade conjunta dos animais expostos e infectados. Exemplares de B. glabrata mineira infectaram-se com cepa simpátrica (MAP) e com 5 alopátricas do Estado de São Paulo (PTH, VPH, OuH, PTR e VPR). B. glabrata paulista mostrou altas taxas de infecção com as cepas MAP, VPR e OuH do trematódeo. Quatro % dos exemplares B. straminea de São Paulo eliminavam cercárias de cepas simpátricas; com cepa mineira apenas 4% apresentaram esporocistos na vigência de 20 miracídios por molusco; as menores taxas de mortalidade foram registradas com essa espécie de molusco, não sendo maior do que 20%. B. tenagophila paulista foi suscetível apenas às linhagens simpáticas sendo 6% a maior taxa de moluscos que eliminaram cercárias. Os resultados indicam que os movimentos populacionais humanos dentro do território paulista e para fora dele são importantes na disseminação da esquistossomose mansônica.

Polymerase chain reaction and restriction fragment length polymorphism analysis of the ITS2 region for differatiation of Brazilian Biomphalaria intermediate hosts of the Schistosoma mansoni

We sequenced the internal transcribed spacer 2 of the ribosomal DNA (ITS2-DNAr) from the three Schistosoma mansoni intermediate hosts in Brazil: Biomphalaria glabrata, Biomphalaria tenagophila and Biomphalaria straminea. Analysis of a restriction map from those sequences allowed us to select putative restriction enzymes able to identify the snail species under study. Four restriction enzymes were used and HpaII provided simple species-specific profiles easily visualized in polyacrylamide gels. The use of ITS2 is advantageous as it provides a small fragment of 460 bp which may be easily amplified by PCR. In the current work, we showed that the amplification of ITS2-DNAr together with HpaII enzyme restriction is an auxiliary molecular tool for the morphological identification of such snails as well as for taxonomic and phylogenetic studies of neotropical planorbids.

The Pampulha reservoir remains a potential urban focus of schistosomiasis mansoni in Brazil: changes in the occurrence patterns of Biomphalaria species and a new record of the parasite

Introduction The Pampulha reservoir has long been a focus of schistosomiasis transmission in Belo Horizonte, State of Minas Gerais, Brazil. The last malacological study conducted in this urban reservoir was more than two decades ago, and thus, an update on the distribution of the species of Biomphalaria as well as new data on the presence of Schistosoma mansoni in this water body are required. Methods The current distribution of Biomphalaria spp. in the Pampulha reservoir and their infection with S. mansoni was evaluated during 55 malacological surveys conducted between 2009 and 2012. Results Biomphalaria straminea displayed a high population density and distribution, and 13.7% (2,233/16,235) of the specimens collected were infected with larval trematodes other than Schistosoma mansoni. Biomphalaria tenagophila and Biomphalaria glabrata, species currently presenting a restricted distribution and small populations, displayed trematode infection rates of 15.2% (98/644) and 13% (83/640), respectively. Thirteen (2%) specimens of B. glabrata were found to be infected with S. mansoni. In addition, a historical review based on previous and new data on the occurrence patterns of Biomphalaria species in this reservoir is presented. Conclusions The results indicate that the Pampulha reservoir remains a potential focus of urban schistosomiasis in Brazil, and significant changes in the occurrence patterns of Biomphalaria species were verified.

Interaction between primary and secondary sporocysts of Schistosoma mansoni and the internal defence system of Biomphalaria resistant and susceptible to the parasite

The outcome of the interaction between Biomphalaria and Schistosoma mansoni depends on the response of the host internal defence system (IDS) and the escape mechanisms of the parasite. The aim of this study was to evaluate the responsiveness of the IDS (haemocytes and soluble haemolymph factors) of resistant and susceptible Biomphalaria tenagophila lineages and Biomphalaria glabrata lineages in the presence of in vitro-transformed primary sporocysts and secondary sporocysts obtained from infected B. glabrata. To do this, we assayed the cellular adhesion index (CAI), analysed viability/mortality, used fluorescent markers to evaluate the tegumental damage and transplanted secondary sporocysts. B. tenagophila Taim was more effective against primary and secondary sporocystes than the susceptible lineage and B. glabrata. Compared with secondary sporocysts exposed to B. tenagophila, primary sporocysts showed a higher CAI, a greater percentage of dead sporocysts and were labelled by lectin from Glycine max and Alexa-Fluor 488 fluorescent probes at a higher rate than the secondary sporocysts. However, the two B. tenagophila lineages showed no cercarial shedding after inoculation with secondary sporocysts. Our hypothesis that secondary sporocysts can escape the B. tenagophila IDS cannot be confirmed by the transplantation experiments. These data suggest that there are additional mechanisms involved in the lower susceptibilty of B. tenagophila to S. mansoni infection.

Focos ativos de esquistossomose mansoni no Vale do Ribeira, Estado de São Paulo, Brasil

São apresentados os resultados das investigações sobre os focos de esquistossomose nas localidades de Pedro de Toledo e Itariri, com referência especial aos índices de infecção natural obtidos para a espécie Biomphalaria tenagophila. Êste molusco, transmissor de esquistossomose no Vale do Ribeira já foi assinalado nos seguintes municípios daquela região: Eldorado, Iguape, Itariri, Jacupiranga, Juquiá, Miracatu, Pariqueraçu, Pedro de Toledo, Registro e Sete Barras. Os índices cercáricos de 68,1%, 75,0% e 81,3% obtidos em alguns lotes de B. tenagophila, os mais altos já verificados para esta espécie, vem mais uma vez comprovar sua grande suscetibilidade à infecção pelo Schistosoma mansoni.

Contribuição ao levantamento da carta planorbídica do Estado de São Paulo: Pesquisa de focos com formas evolutivas do Schistosoma mansoni. II. Município de Peruibe (Litoral Sul do Estado de São Paulo)

Em levantamento realizado em Peruibe, em janeiro de 1966, foram identificados focos de Biomphalaria tenagophila infestados com formas evolutivas do Schistosoma mansoni.

Levantamentos parasitológicos, em particular a esquistossomose mansônica, nas cidades de Itanhaém e Mongaguá (Litoral Sul do Estado de São Paulo)

São apresentados os resultados dos exames parasitológicos obtidos em levantamentos realizados na cidade de Itanhaém, em 1968, e de Mongaguá, em 1969. Entre os casos de esquistossomose registrados não houve nenhum identificado como autóctone das áreas estudadas; os exemplares de Biomphalaria tenagophila coletados estavam negativos para formas evolutivas do Schistosoma mansoni.

Aspectos epidemiológicos da esquistossomose mansônica na região da Represa de Americana, estado de São Paulo, Brasil

Após conhecimento de casos humanos de esquistossomose mansônica supostamente autóctones da Represa de Americana (São Paulo, Brasil) procedeu-se ao estudo epidemiológico da região. Foram constatados seis focos localizados junto à Represa. Foram encontradas as seguintes espécies de moluscos: Biomphalaria tenagophila; B. straminea; B. peregrina; Drepanotrema cimex; D. lucidum; |Lymnaeidae; Ancylidae e Physidae. Exemplares de Biomphalaria tenagophila coletados nos focos apresentaram índices de infecção para cercárias de S. mansoni que variaram de 0,9 a 45%. Mus musculus albinos foram infectados com cercárias no laboratório e nos focos, sendo reproduzido o ciclo do S. mansoni em ambas as condições. Foram registrados 82 casos humanos autóctones de esquistossomose mansônica, na região da Represa de Americana.

Alguns dados sobre o comportamento parasitológico das linhagens humana e silvestre do Schistosoma mansoni, no Vale do Rio Paraíba do Sul, SP (Brasil)

Verificou-se ser elevada a percentagem de animais parasitados por S. mansoni em roedores silvestres, de diferentes gêneros e espécies, capturados no Vale do Rio Paraíba do Sul, SP, Brasil onde se encontram com freqüência, casos humanos de esquistossomose mansônica. A partir de fígados de roedores naturalmente infectados foram obtidos miracídios para o isolamento da linhagem silvestre (S). Para o isolamento da linhagem humana (H), foram utilizados miracídios procedentes de fezes de doentes comprovadamente autóctones do Vale do Rio Paraíba. Estudou-se, comparativamente, o comportamento das duas linhagens em B. tenagophila que é o hospedeiro intermediário natural na região e em camundongo albino, utilizado como hospedeiro definitivo. Verificou-se que: a taxa de mortalidade de B. tenagophila infectadas com a linhagem "S" não é, estatisticamente, diferente da dos moluscos utilizados para controle; é significativa a diferença entre a taxa de mortalidade verificada nos moluscos infectados com a linhagem "S" e com a linhagem "H", a qual é consideravelmente maior nos moluscos infectados com a linhagem "H"; nos camundongos infectados com a linhagem "H" verificou-se ser significativo o coeficiente de correlação entre o número de granulomas hepáticos e o de trematódeos adultos; nos camundongos infectados com a linhagem ''S" não houve possibilidade do estabelecimento de tal correleção.

Infecção natural de pequenos mamíferos por Schistosoma mansoni, na represa de Americana (São Paulo, Brasil)

No período de janeiro de 1972 a agosto de 1975, nas margens da Represa de Americana (= Represa de Salto Grande), Brasil, em locais onde foi constatada a infecção por Schistosoma mansoni no homem e em Biomphalaria tenagophila, foram capturados 124 pequenos mamíferos: 44 exemplares de Cavia aperea aperea; 9 de Didelphis albiventris; 7 de Holochilus brasiliensis leucogaster; 39 de Lutreolina crassicaudata; 3 de Mus musculus brevirostris; 14 de Oryzomys nigripes eliurus; 1 de Oryzomys subflavus; 4 de Rattus rattus e 3 de Zygodontomys brachyurus. Verificou-se a infecção natural por S. mansoni em 2 exemplares de D. albiventris, em 3 de H. b. leucogaster e em 5 de L. crassicaudata. Estudou-se nestes animais a presença de elementos esquistossomóticos nas fezes e no intestino, bem como a distribuição e sexo de vermes adultos no sistema porta. Nas fezes de L. crassicuadata e de D. albiventris raramente foram observados ovos maduros do trematódeo. Em H. b. leucogaster era comum a presença destes elementos nas fezes. Devido a pequena abundância de H. b. leucogaster na área estudada, é provável que este roedor pouco contribua na disseminação de ovos de S. mansoni. Parece que a fauna de pequenos mamíferos estudada nesta região não desempenha papel relevante na manutenção do ciclo de S. mansoni.