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.

Hybridism between Biomphalaria cousini and Biomphalaria amazonica and its susceptibility to Schistosoma mansoni

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.

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.

The relationship between genetic variability and the susceptibility of Biomphalaria alexandrina snails to Schistosoma mansoni infection

In the present study, Biomphalaria snails collected from five Egyptian governorates (Giza, Fayoum, Kafr El-Sheikh, Ismailia and Damietta), as well as reference control Biomphalaria alexandrina snails from the Schistosome Biological Supply Center (SBSC) (Theodor Bilharz Research Institute, Egypt), were subjected to species-specific polymerase chain reaction (PCR) assays to identify the collected species. All of the collected snails were found to be B. alexandrina and there was no evidence of the presence of Biomphalaria glabrata. Randomly amplified polymorphic DNA (RAPD)-PCR assays showed different fingerprints with varying numbers of bands for the first generation (F1) of B. alexandrina snail populations (SBSC, Giza, Fayoum, Kafr El-Sheikh, Ismailia and Damietta). The primer OPA-1 produced the highest level of polymorphism and amplified the greatest number of specific bands. The estimated similarity coefficients among the B. alexandrina populations based on the RAPD-PCR profiles ranged from 0.56 (between SBSC and Ismailia snails) to 0.72 (between Ismailia and Kafr El-Sheikh snails). Experimental infection of the F1 of progeny from the collected snails with Schistosoma mansoni (SBSC strain) showed variable susceptibility rates ranging from 15% in the Fayoum snail group to 50.3% in SBSC snails. A negative correlation was observed between the infection rates in the different snail groups and the distances separating their corresponding governorates from the parasite source. The infection rates of the snail groups and their similarity coefficients with SBSC B. alexandrina snails were positively correlated. The variations in the rates of infection of different B. alexandrina groups with S. mansoni, as well as the differences in the similarity coefficients among these snails, are dependent not only on the geographical distribution of the snails and the parasite, but also on the genetic variability of the snails. Introduction of this variability into endemic areas may reduce the ability of the parasite to infect local hosts and consequently reduce schistosomiasis epidemiology.

The significance of the amoebocyte-producing organ in Biomphalaria glabrata

In molluscs, internal defence against microorganisms is performed by a single cell type, i.e., the haemocyte or amoebocyte. The origin of these cells in Biomphalaria glabrata was initially thought to be localised within the vasculo-connective tissue. More recently, origin from a single organ, termed the amoebocyte-producing organ (APO), has been postulated based on the occurrence of hyperplasia and mitoses during Schistosoma mansoni infection. The present investigation represents a histological, immuno-histochemical and ultra-structural study of the B. glabrata APO, whereby histological identification was facilitated by means of collecting epithelial basophilic cells. These cells were comprised of single-cell layers that cover a portion of the stroma, which contains many small, round cells and haemolymph sinuses, as well as a small area of the pericardial surface of the reno-pericardial region. On occasion, this epithelial component vaguely resembled the vertebrate juxtaglomerular apparatus, which reinforces its presumed relationship to the kidney. Both in normal and infected molluscs, mitoses were only occasionally found. The present quantitative studies failed to demonstrate the presence of APO cellular hyperplasia, either in normal or schistosome-infected B. glabrata. Conversely, several structural details from the APO region in B. glabrata were found to be consistent with the hypothesis that the APO is a filtration organ, i.e., it is more closely related to the kidney rather than the bone marrow, as has been suggested in the literature.

Aspectos físico-químicos e biológicos relacionados à ocorrência de Biomphalaria glabrata em focos litorâneos da esquistossomose em Pernambuco

This study analyzes the physico-chemical and biological parameters of freshwater aquatic environments in coastal areas of the State of Pernambuco, Brazil, where snail vector foci and cases of schistosomiasis have been recorded. The tests were carried out using standard methods and sediments were analyzed using the X-ray fluorescence technique. The micro-algae were analyzed using conventional microscopy. The most abundant chemical elements in the sediments were Fe, Ca, Si and Al and there were high concentrations of electrolytes in comparison with data published in the literature. The mean values for salinity (7.7 g/L) and sulfate (489 mg/L) were much higher than normal for freshwater environments. The micro-algae identified are those that inhabit environments with higher levels of salinity. The snail Biomphalaria glabrata seems, therefore, to be more tolerant to salty environments and this constitutes a risk of spread of schistosomiasis.

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.

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.

Alimentação e fecundidade de planorbídeos criados em laboratórios: IV - Helisoma duryi (Wetherby, 1879). (Pulmonata, Planorbidae)

Utilizando dois tipos distintos de alimentos: "Aquariol" (alimento utilizado para alimentar peixes ornamentais) e alface, isolados e associados, na criação de Helisoma (Seminolina) duryi (Wetherby, 1879) em laboratório, foi verificado que; 1. Os caramujos alimentados com alface fresca liberaram, em 17 semanas, 826 desovas on 17.707 ovos. 2. Os especimens alimentados com "aquariol" depuseram 534 desovas on 11.361 ovos. 3. O oferecimento dos dois materiais associados resultou no aumento da fertilidade, obtendo-se, assim, 1.355 desovas ou 34.645 ovos. 4. Em termos médios, cada conjunto de quinze exemplares (com 10 a 14 mm de diâmetro) liberou, diariamente, 95,5 - 148,8 e 291,1 ovos/dia, segundo o tipo de alimento consumido: "aquariol", alface fresca, ou ambos associados. O número de desovas/dia, na mesma ordem enunciada, foi de 4,5 - 6,9 e 11,4, respectivamente. 5, Os exemplares utilizados descendiam de uma cepa supostamente recém introduzida no município de Formosa, GO (Brasil)e originária da Região Neártica (EUA). Foi comentada a possibilidade de controle biológico de diferentes espécies de Biomphalaria através de H. duryi - questão sobre a qual vêm desenvolvendo investigações biológicas e ecológicas.

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.