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.

Nodulação e micorrização em Anadenanthera peregrina var. falcata em solo de cerrado autoclavado e não autoclavado

Anadenanthera peregrina var. falcata (angico-do-cerrado), uma leguminosa arbórea, forma associações simbióticas com bactérias fixadoras de nitrogênio (rizóbios) e com fungos micorrízicos arbusculares. Com o objetivo de avaliar a eficiência da inoculação de fungos micorrízicos e rizóbios no crescimento inicial de plantas de angico-do-cerrado, crescidas em solo autoclavado e em solo não autoclavado com e sem inoculação, foi desenvolvido um experimento em casa de vegetação, utilizando raízes micorrizadas de milho e uma mistura de isolados de rizóbios como inoculantes. O crescimento das plantas foi influenciado positivamente pela concomitante inoculação do fungo micorrízico e do rizóbio, tendo as plantas desse tratamento apresentado biomassa cerca de 60 % maior do que o controle no décimo mês. A inoculação de apenas um dos microssimbiontes, entretanto, não provocou diferença na produção de biomassa das plantas. A percentagem de colonização micorrízica foi significativamente mais alta e o número de nódulos maior nas raízes das plantas crescidas no solo não autoclavado, ocasionados pela população de fungos e rizóbios nativos. Nesse tratamento, houve pequeno acúmulo de matéria no xilopódio, provavelmente em virtude do dreno fotossintético por parte dos microssimbiontes, e a concentração de P na parte aérea e xilopódio dessas plantas foi cerca de 1,2 e 8 vezes maior, respectivamente, por causa da colonização micorrízica.

Estudo fitoquímico e avaliação alelopática de Memora peregrina - "ciganinha" - Bignoniaceae, uma espécie invasora de pastagens em Mato Grosso do Sul

Memora peregrina (local name: 'ciganinha' - Bignoniaceae) is a weed that often invades pastures in the Brazilian state of Mato Grosso do Sul. From its leaves and subterranean parts, the following compounds were isolated: allantoin (20.7 w/w in subterranean parts), the iridoid 6beta-hydroxyipolamiide, hyperin, 3'-O-methylhyperin, 4-hydroxy-N-methylproline, beta-sitosterol, alpha-amirin and beta-amirin, and lupeol. Allantoin exhibited an activity of inducing germination in seeds of Lactuca sativa used as a biological model, and the iridoid showed moderate activity in the larval development of Anagasta kuehniella. These results, associated with this plant's behavior, are suggestive of the occurrence of adaptive and competitive strategies in relation to other plant species.

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.

Parâmetros morfológicos de mudas de Sesbania virgata (Caz.) Pers e de Anadenanthera peregrina (L.) cultivadas em substrato fertilizado com composto de lixo urbano

Objetivou-se avaliar o efeito do composto de lixo urbano nos parâmetros morfológicos de mudas de sesbânia e de angico. Os substratos de cultivo tiveram as seguintes proporções de composto e de amostras de subsolo de Neossolo Quartzarênico ou de Latossolo Vermelho-Amarelo (%): 0:100; 20:80; 40:60; 60:40 e 80:20, sem a utilização de fertilização mineral. As sementes de sesbânia foram inoculadas com a estirpe recomendada BR 5401. Os parâmetros morfológicos das mudas de sesbânia e angico, suas relações e índice de qualidade de Dickson (IQD) foram determinados aos 56 e 120 dias após a semeadura, respectivamente para cada espécie. Os efeitos dos substratos nos parâmetros morfológicos das mudas de sesbânia e angico variaram de acordo com as proporções do substrato. Para o angico, a adição do composto proporcionou aumento na altura da parte aérea, razão entre massa seca de parte aérea por massa seca de raiz e IQD. Para sesbânia, a adição do composto de lixo ao substrato proporcionou aumento na altura da parte aérea, diâmetro do coleto, massa seca de raiz, da parte aérea e total, razão entre massa seca da parte aérea por massa seca de raiz, IQD e número de nódulos. A obtenção de máxima produção de matéria seca total, diâmetro do coleto e IQD foi de 57:43, sendo, portanto, esta a proporção composta de lixo:solo recomendada para a produção de mudas de sesbânia.

Evaluation of the physiological quality of anadenenthera peregrina (L.) Speg. seeds during storage

The aim of this experiment was to evaluate the effect of storage temperature on the viability and vigour of A. peregrina seeds. Seeds of A. peregrina harvested in Viçosa-MG, in September 2005 were used. The seeds were stored in the Laboratory of Forest Seeds Analysis (LASF) of the Universidade Federal de Viçosa (UFV), in a cold chamber, under approximately 5ºC over150 days. The seeds were stored under 20ºC for 150 days more. Seed moisture was determined after oven-drying at 105 ± 3ºC for 24 hours. The seeds were analyzed monthly using germination, tetrazolium and electrical conductivity tests and Osmopriming during 300 days, with four repetitions of 25 seeds. A. peregrina seeds kept a constant percentage of germination and viable seeds under the tetrazolium test for five months of storage, under either 5ºC and 20ºC. The electrical conductivity test showed significant differences in relation to the storage period, proving to be more sensitive than the standard germination test. Seed priming in PEG -0,4 MPa solution was not efficient in recovering the viability and vigor of the stored seeds at 5ºC and 20ºC.

Propriedades de chapas de aglomerado fabricadas com adesivo tânico de angico-vermelho (Anadenanthera peregrina) e uréia-formadeído

O principal objetivo deste trabalho foi produzir chapas de aglomerado substituindo-se parcial ou totalmente os adesivos à base de uréia-formaldeído por adesivos de taninos extraídos da casca de angico-vermelho (Anadenanthera peregrina) e hidrolisados. As chapas de aglomerado foram fabricadas com partículas de Pinus elliottii e dimensões aproximadas de 40 x 40 x 1 cm, contendo 8% de sólidos de adesivo em relação à massa seca de partículas, sendo 100% de adesivo de uréia-formaldeído puro ou contendo 12,5, 25 e 37,5% de adesivo tânico. Foram produzidas, também, chapas com adesivos tânicos puros, constituindo um total de cinco tratamentos, com três repetições. As chapas foram fabricadas com densidade média de 0,7 g/cm³. As propriedades físicas e mecânicas das chapas foram determinadas em conformidade com a norma ASTM D-1037 (1993). Os resultados dos testes mecânicos foram comparados com os valores mínimos estabelecidos na norma ANSI/A 208.1-1993 (Wood Particleboard), enquanto os resultados dos testes de absorção de água e inchamento em espessura, comparados com os valores máximos estabelecidos na norma DIN 68m761 (1)-1961. A resistência mecânica das chapas de aglomerado produzidas com adesivos contendo taninos de angico-vermelho, com exceção do módulo de elasticidade, ultrapassou os valores mínimos requeridos na norma ANSI/A1-280/93. A absorção de água e o inchamento em espessura, após 2 e 24 h de imersão, de todas as chapas excederam o valor máximo estabelecido. Conclui-se que os adesivos de taninos de angico-vermelho podem substituir parcial ou totalmente o adesivo comercial de uréia-formaldeído, e as suas misturas são indicadas para uso interior, em que a resistência à umidade não é requerida.

Avaliação da viabilidade e vigor de sementes de Anadenanthera peregrina (L.) Speg. submetidas ao envelhecimento acelerado e ao osmocondicionamento

O objetivo deste trabalho foi avaliar o vigor e viabilidade de sementes de Anadenanthera peregina submetidas ao envelhecimento acelerado e ao osmocondicionamento. As sementes foram acondicionadas em caixas tipo "gerbox" com uma tela de alumínio separando-as do fundo do gerbox, onde se adicionou água destilada. As caixas foram mantidas em câmara de envelhecimento nas temperaturas de 40, 50 e 60 ºC, durante 24, 48, 72 e 96 h. Para o controle foram utilizadas sementes não submetidas ao envelhecimento. Após cada tempo, as sementes foram submetidas aos seguintes testes: teste de germinação, índice de velocidade de germinação, condutividade elétrica, teste de tetrazólio, osmocondicionamento e avaliação das plântulas. A 40 ºC, a porcentagem de germinação, o Índice de Velocidade de Germinação (IVG) e a porcentagem de sementes viáveis pelo teste de tetrazólio decresceram significativamente. Houve incremento significativo da curva de condutividade elétrica à medida que aumentou a permanência das sementes dentro da câmara de envelhecimento. A partir de 96 h de permanência na câmara de envelhecimento, as sementes perderam toda a viabilidade, assim como aquelas submetidas ao envelhecimento a 50 e 60 ºC. Na avaliação das plântulas, todos os parâmetros analisados decresceram à medida que as sementes permaneceram na câmara de envelhecimento por mais tempo, com exceção da porcentagem de plântulas anormais e do peso fresco da raiz, que tiveram incremento nos seus valores.

Parâmetros cinéticos de adesivos produzidos a partir de taninos de Anadenanthera peregrina e Eucalyptus grandis

Devido à crise do petróleo, na década de 70, pesquisas tiveram o objetivo de substituir parcial ou totalmente os adesivos convencionais, à base de petróleo. Uma forma de substituição foi a utilização de fontes naturais, como os taninos, provenientes de algumas espécies arbóreas. Porém, os taninos apresentam algumas limitações de uso, que podem ser superadas através da sulfitação e da hidrolise ácida. Portanto, o objetivo deste trabalho foi avaliar o efeito da hidrólise ácida e sulfitação de taninos extraídos da casca de Eucalyptus grandis e Anadenanthera peregrina, nos parâmetros cinéticos dos adesivos tânicos, através da técnica de calorimetria diferencial exploratória (DSC). Os taninos foram hidrolisados com ácido acético ou ácido clorídrico a 10N, empregando-se três valores de pH e três tempos de reação. A sulfitação foi realizada em solução de sulfito de sódio, à 5%, em três tempos de reação. A energia de ativação, entalpia e temperatura de cura dos adesivos foram reduzidas pela hidrólise e sulfitação dos taninos, porém apresentaram banda larga de polimerização. O tempo de reação dos taninos não teve efeito sobre os parâmetros cinéticos.

Anatomical and ultrastructural aspects of root and mycorrhiza of Anadenanthera peregrina (L.) Speg. var. falcata (Benth.) Altschul (Leguminosae-Mimosoideae)

A. peregrina var. falcata form mutualistic symbiosis with arbuscular mycorrhizal fungus. An anatomical and ultrastructural study was carried out to analyze some aspects of this simbiotic association as well as some root features. The results evidenced the presence of fibers with non-lignified thicked secondary walls in the stele and sparse papillae on root surface. A. peregrina var. falcata mycorrhizas presented features of Arum-type (intercellular hyphae) and Paris-type (extensive coils) arbuscular mycorrhiza. Their general appearance with extraradical hyphae, intracellular coils, intercellular hyphae and arbuscules, is in agreement with arbuscular mycorrhizas of several plants. The ultrastructural observations showed that in intercellular hyphae and arbuscules vacuoles were dominant and that in rough endoplasmatic reticulum and small vesicles seems to be associated with arbuscule senescence process.

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.